18th annual midwest swine nutrition conference proceedings€¦ · cj america cooper farms csa...

18th AnnualMidwestSwineNutrition ConferenceProceedings

Indianapolis, Indiana—September 6, 2018

Midwest Swine Nutrition Conference

2018 SponsorsAB Vista Feed Ingredients

ADM Animal NutritionAgri-King

Ajinomoto Animal NutritionAlltech

APC CompanyBASF Corporation

Cargill Animal NutritionChr. Hansen Animal Health and Nutrition

CJ AmericaCooper Farms

CSA Animal NutritionDarling Ingredients

Distributors ProcessingDSM Nutritional Products

DuPont/Danisco Animal NutritionElanco Animal Health

Evonik-Degussa CorporationFats and Proteins Research Foundation

Hubbard FeedsInternational Ingredient Corporation

Kalmbach FeedsKemin

Kent Nutrition GroupKing Technica Group

Lallemand Animal NutritionMicronutrients

MonsantoNational Pork Board

Norel Animal NutritionNovus International

NutriadNutriQuest

Phibro Animal HealthPhileo Lesaffre Animal Care

PIC North AmericaProvimi North America

Purina Animal NutritionRalco NutritionStuart ProductsThe Maschhoffs

United Animal HealthVita Plus CorporationZinpro Corporation

Zoetis

Appreciation is expressed to the Indiana Farm Bureau and their staff for hosting the 2018 Midwest Swine Nutrition Conference and providing the facilities for this function for the past 14 years.

Industry RepresentativesDennis Liptrap, Ralco Nutrition—Overall Chairman

Ronnie Moser, United Animal HealthCasey Bradley, DSM Nutritional Products

University of KentuckyGary Cromwell—Sponsor Recruitment, Treasurer, Editor of Proceedings

Merlin LindemannPurdue University

Scott Radcliffe—VideographerBrian Richert

Tip ClineUniversity of Illinois

Hans SteinJim PettigrewRyan Dilger

Michigan State UniversityDale Rozeboom

Gretchen HillThe Ohio State University

Sheila Jacoby

Setup and Printing of ProceedingsDennis Duross, Ag Communications Services,

University of Kentucky

Website Development and MaintenanceCody Ortt and Velvet Barnett

University of Kentucky

Meeting Room and FacilitiesIndiana Farm Bureau

Randy Kron, PresidentDiane Helton, Administrative Assistant

Lunch and Refreshments at BreaksAramark

Mario Puccinelli, Food Service Director


Program Committee


Schedule of Presentations

8:15 a.m Registration

9:00 Welcome and IntroductionsDennis Liptrap, Ralco Animal Nutrition

9:05 The State of the U.S. Animal Protein SectorPablo Sherwell, Head of Rabo Research Food and Agribusiness NA, Rabobank

9:50 Potential Nutritional Considerations to Prevent Sow MortalityCasey Bradley, DSM Nutritional Products and Monique Pairis-Garcia, The Ohio State University

10:25 Break

11:00 Impact of Dietary Amino Acid Balance on Nitrogen and Energy Efficiency in Sows: From Theory to PracticeNathalie Trottier, Michigan State University

11:30 Copper Levels and Sources for SowsMerlin Lindemann, University of Kentucky

12:00 Lunch

1:00 Development and Nutritional Value of Advanced Soybean Products Used in Diets for Young PigsHans H. Stein, University of Illinois

1:35 Nutritional Regulation of Piglet Gut HealthSheila Jacobi, The Ohio State University

2:10 Break

2:40 Glutamine and Transport StressBrian Richert, Purdue University

3:15 Strategic Therapeutic Antibiotic Use Compared to the Challengeof Not Using Antibiotics for Growing PigsDean Boyd, The Hanor Company


Table of Contents

The State of the U.S. Animal Protein Sector ................................................................................5Pablo Sherwell

Potential Nutritional Considerations to Prevent Sow Mortality .........................................7Casey L. Bradley, Jon Bergstrom, and Monique Pairis-Garcia

Impact of Dietary Amino Acid Balance on Nitrogen and Energy Efficiency in Sows: from Theory to Practice .............................................................. 15Trottier, N. L., S. Zhang, and J. S. Johnson Copper Levels and Sources for Sows ........................................................................................... 21Ning Lu and Merlin Lindemann Development and Nutritional Value of Advanced Soybean Products Used in Diets for Young Pigs .......................................................................................................... 29Laia Blavi and Hans H. Stein

Nutritional Regulation of Piglet Gut Health ............................................................................. 33Sheila K. Jacobi, Emily E. Chucta, Ariel Taylor, and Jack Odle Glutamine and Transport Stress ................................................................................................... 39Alan Duttlinger, Brian Richert, Donald C. Lay Jr., and Jay S. Johnson Strategic Therapeutic Antibiotic Use Compared to the Challenge of Not Using Antibiotics for Growing Pigs................................................... 51R. Dean Boyd, Tara S. Donovan, and Cate E. Rush

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The State of the U.S. Animal Protein Sector

Pablo Sherwell RaboResearch

New York Phone: 212-808-6822

[email protected]

The U.S. animal protein sector is going through an inflection point. After a few years of posi-tive margins, the sector is moving to an economic downturn. Hogs, in particular, are shifting to the negative side, and this scenario is likely to deteriorate faster and deeper than previously anticipated. This inflection point in the industry comes after several subsectors began investing and ex-panding around the same time in 2015/16. This expansion is the natural result of stable domestic meat consumption combined with strong exports, positive margins, and limited production ca-pacity However, the economic scenario for the sector is changing as there are many factors that came into play concurrently. First, the expansion is tangible in almost every protein (except for turkey), and it is creating an oversupply and, consequently, more competition among meats. This year, beef and pork pro-duction will grow over 4 percent respectively, poultry only around 1.5 percent, but all in all meat production will increase around 3.6 percent. In addition, plans to keep expanding in 2019 are (at least at this point) on their way. Second, new packing plants are facing tremendous challenges securing adequate labor. The boost in the economy and the increase in labor demand in other industries, such as energy, con-struction, restaurants, etc., is leaving the meat industry with few options. Changes in demograph-ics and immigration laws have only made the situation worse. Third, a trade war with two of the most important U.S. meat customers, Mexico and China, is complicating the allocation of additional supplies. Moreover, since there is not a timeline on how long this is going to last, planning uncertainty is rising. As long as these tariffs are in place, other competitors such as Brazil, Europe, and Canada will gain market share in traditional U.S. export markets. As a result of the above, we anticipate animal protein margins to keep declining until a more balanced market is reached. However, a correction in the market will take time as investments in the industry have only recently been made. In fact, we expect that margins will deteriorate further before they recover. The situation for packers could be more controllable as long as they are able to manage margins. The presentation will conclude with our mid-term outlook for the sector and discuss some challenges and opportunities as we move forward.

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Potential Nutritional Considerations to Prevent Sow MortalityCasey L. Bradley, Ph.D., and Jon Bergstrom, Ph.D.

DSM Nutritional Products, NA 45 Waterview Boulevard

Parsippany, New Jersey 07054 Phone: 479-366-8488

[email protected]

Monique Pairis-Garcia, DVM, Ph.D. The Ohio State University

Department of Animal Science Columbus, OH

Phone: 951-442-0429 [email protected]

SummarySow mortality has always been a challenge for the swine industry, but there has been a spike in sow mortality over the last few years, with limited understanding of the root causes. Furthermore, the incidences of sow uterine and rectal prolapses have also substantially increased within this time frame. It is well documented that lameness is also a leading cause of mortality, and most likely the predominant or secondary reason for culling in sow herds.

Within human medicine, Pelvic Floor Disorder and/or vaginal prolapses are also anticipated to steadily increase in next few decades; thus, there is substantial research dedicated to understand-ing this similar issue we are observing in sows with prolapses. The main characteristics identified in human subjects are impaired collagen and elastin metabolism in the supporting ligaments and muscle architecture. Furthermore, metabolic stressors such as mycotoxin contaminated feeds and/or oxidative stress can alter nutrient requirements of animals and potential hormonal responses within the sow. For example, zearalenone is known to have estrogen like responses that could cause laxity issues within ligaments like that found in human medicine, resulting in injury and thus lame-ness or uterine ligament support.

Vitamin C is an essential component in collagen and elastin metabolism, but a dietary requirement has not been established for swine because they are generally considered capable of synthesizing enough on their own. Additionally, there is a considerable amount of data indicating that the greater bioavailability of certain trace minerals, especially organic vs. inorganic sources, may reduce lame-ness and mortality in sow herds. In humans and swine, there is mounting evidence that glycine, a “nonessential” amino acid, is likely deficient for optimizing certain metabolic pathways, including collagen formation, especially during pregnancy. Thus, the objective of this paper is to present novel perspectives from human medicine, and discuss the potential for nutrients that are often overlooked in today’s nutritional programs for sows.

Introduction Sow mortality has always been a concern in sow herds, and any percentage of sow mortality presents challenges for all management levels. It is well known that replacement gilts do not start becoming profitable until their third or fourth parity within a system. Thus,

increasing discussions around rising sow mortality rates in the USA over the last years has involved nearly all segments of the industry. Nevertheless, the real ques-tion remains largely unanswered: What are the primary underlying causes for the apparent increase in sow mor-tality and, ultimately, what can we focus on with inter-ventions?

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Hindering our ability to define strategies to combat mortality, there is limited information on the precise reasons that sows are dying. This is largely due to impre-cise record keeping and the lack of expertise and time to conduct necropsies on farms. In 2007, Sanz et al. as-sessed sow mortality in a 5,200-sow unit over a 20-wk period in which all deaths were necropsied. The authors reported that 47% of sows were euthanized, 28% were expected deaths, and 25% died unexpectedly. Locomo-tion issues were the main reason for sow mortality in all stages of production, with arthritis and osteochondro-sis-dyschondroplasia as the main lesions found. Other reasons for mortality included: prolapses, farrowing difficulty, gastric ulcers, and circulatory or respiratory problems. Most recently, Dr. Jeremy Pittman of Smithfield Foods discussed the rise in mortality in sows (as re-ported by Day, 2017) and the dramatic increase in pro-lapses. He also reported a long list of potential factors: mycotoxins, hypocalcemia, vitamin deficiency, use of phytase, litter sizes, constipation, genetics, change in estrogen and/or other hormone levels, oxidative stress, meal vs. pelleted feed, high and rapid feed intake, bloat-ing, coughing, piling, housing design, diarrhea, colitis, and excess lysine. Without any clear solutions to the current mortality problems, the objective of this paper is to discuss potential similarities with human medicine and the possibilities for nutritional interventions.

Uterine Prolapses—Human Medicine Perspective Pelvic Floor Disorder and vaginal and/or uterus prolapses have also been a problem in human medicine. Multiple-pregnancies are known as one of the main cor-relating factors for these issues in women; however, with modern advances in science, these problems have been studied in greater detail. Even in humans, the incidence of these cases continues to increase, and are estimated to increase by 45% or double in frequency over the next 30 years, which may be related to the aging population (Luber et al, 2001).

Swine as a Biological Model for Humans There are two main supportive tissues for the repro-ductive tract, supportive ligaments and muscles. The pelvic support ligaments, cardinal and uterosacral liga-ments (Figure 1), suspend the pelvic organs to the pelvic side walls over the levator plate, while the pelvic floor muscles close the urogenital hiatus, allowing the organs (uterus, bladder, rectum) to rest on top of them. Work conducted at Virginia Tech University, Tan (2015) dis-sected the tissues of sows to better understand the me-

chanical properties of the supportive ligaments and how they would compare to humans for use as a potential biological model. He reported finding loose connective tissue in all specimens, with a larger amount of ground substance in the left cardinal ligament (LCL), while the right cardinal ligament (RCL) contained a larger num-ber of smooth muscle fibers and adipose cells. Within the uterosacral ligament (USL) there was dense con-nective tissue, characterized by considerable amounts of collage and elastin fibers, but this was not identified in the other ligaments. Additionally, elastin content was significantly higher in the USL than either CL. Tensile strength was found to vary considerably with the loca-tion of the ligaments, but was relatively similar within the same ligament. As noted with Tan’s work, collagen and elastin are the two main structural constituents of supportive ligaments. During pregnancy, elastin within the female reproduc-tive organs (which provides extension and recoil to tis-sues) will dramatically increase and then decrease back to pre-gravid levels after parturition. Within the litera-ture on humans, there are several publications indicating that improper collagen and elastin metabolism is corre-lated to prolapses or pelvic floor disorders in women. For instance, Zong et al. (2010) reported that tropoelastin

Figure 1. Adapted from Tan et al., 2015. A. Sow reproductive support ligaments: Left cardinal ligament (LCL), uterosacral ligament (USL), and right cardinal ligament (RCL) and their lo-cation relative to the rectum or vagina. B. Ligament (LCL, USL, and RCL) attachment to the cervix.

B.

A.

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(432%), mature elastin (55%), proMMP-9 (90%), and ac-tive MMP-9 (106%) were increased in women with pro-lapses, while active MMP-2 (41%) was decreased when compared to the control. Furthermore, Megadhana and Suwiyoga (2016), found high expression of estrogen re-ceptor alpha and collagen III with low expression of elas-tin in sacrouterine ligaments as risk factors in stage III-IV uterine prolapses. Because of the change in collagen or elastin metabolism, the tissue that is formed is weak, but may not be less, as Hahn et al. (2014) reported an in-creased diameter of the collagen fibril that lacked proper formation and strength for women with a pelvic organ prolapse when compared to the control.

Hormonal Influences In a review, Leblanc et al. (2017) discussed the importance of estrogens as a regulating factor of me-tabolism for connective tissues, like bone, muscle, and cartilage. Primarily they influence the development, maturation, and function of the female reproductive or-gans, but are also involved in developmental processes of the bone formation. A great example of how estro-gen may impact mobility or lameness can be shown in humans, in which injury rate of the cruciate ligament is approximately 2 to 8 times higher than in men. Further-more, pre-menopausal women are at greater risk, and there is some evidence that suggests tendon-injuries in female athletes might differ in different phases of the menstrual cycle, with the laxity in the knee of women peaking between ovulation and post-ovulation. Addi-tionally, the authors discussed the change in “stiffness” of a tendon; whereas, relaxin, for example, decreases the stiffness of some ligaments during pregnancy and leaves the female open to specific injuries. Furthermore, Vitamin D is a steriod prohormone that is essentially in-volved in regulating calcium homeostasis, but may also be important in regulating reproductive processes by influencing estrogen synthesis (Reese and Casey, 2015).

Collagen and Osteochondrosis It has been noted that many lame and culled sows have osteochondrosis (OC) lesions (Figure 2) in their joints at slaughter, and that this is also a leading reason for lameness in sows. It has been suggested that OC lesions, or the development of lesions, can potentially arise in utero. Early work has defined OC as focal failure of endochondral ossification with subsequent persis-tence of the growth cartilage in the epiphyseal growth plates. These lesions can heal over time, but scar tissue can still be seen in older animals. Genetics, conforma-tion, injury or trauma, and nutritional deficiencies have all been linked to the prevalence of OC. However, re-

cently Laverty and Girad (2016) indicated that there are now several studies demonstrating a consistent increase in type II collagen synthesis in young animals of mul-tiple species. This increase in type II collagen synthesis may be indicative of repair, but there is a lack of coor-dinating synthesis of the cartilage matrix proteoglycan molecule. This appears to demonstrate altered type II collagen metabolism may be involved in early stages of OC; thus, indicating an importance of collagen metabo-lism for both bone and ligaments in swine.

Other Potential Metabolic Stressors Mycotoxins Zearalenone is the primary mycotoxin that comes into question when we are having reproductive issues in sows. However, many times feed samples are obtained and no mycotoxins are present, or low levels are found and not considered problematic. Nevertheless, there is a growing body of evidence that zearalenone concen-trations as low as 75 ppb can impact the reproductive tract in as little as 6 days post contamination. Research conducted with 10.8 kg gilts indicated that both length and weight of the reproductive tract was increased with zearalenone contaminated feed (Oguey and Du-rox, 2016). This is alarming because, traditionally, gilts of this size are fed a standard nursery program and not

Figure 2. Osetochondrosis lesion in a femur of a growing pig (photo provided by T. Crenshaw, University of Wisconsin, Madison).

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a specialized gilt development program. There is, how-ever, limited research evaluating whether pulses of con-taminated feed throughout a gilt or sow’s lifetime may impact reproductive health and longevity. Additionally, mycotoxin mitigation could potentially interfere with the absorption of certain essential vitamins and other nutrients, and that may or not be considered during diet formulation.

Lameness and Oxidative Stress Within dairy, it has been demonstrated that oxida-tive stress is involved in lameness. For instance, Zhoa et al. (2015) reported that lame cows had a 17.7% increase in malondialdehyde (MDA), a 7.7% decrease in superox-ide dismutase (SOD), 18% decrease in metallothionein (MT), and 16% increase in the oxidized glutathione to reduced glutathione (GSSG/GSH) ratio. Furthermore, trace mineral metabolism was altered as indicated by the reduction of trace mineral levels (Zn, Cu, Mn) in serum, hair, and hoof keratin, where these minerals are known to play a role in protection from oxidative stress. Furthermore, CTX-II and COMP was elevated in the serum lame cows, indicative of cartilage breakdown.

Micronutrient ConsiderationsVitamin C It has generally been considered that swine can en-dogenously produce their own Vitamin C to meet their requirements, and that they may only require supple-mentation during stressful conditions. However, a line of pigs has been identified to have a mutated gene so that they lack the ability to synthesize Vitamin C. This genetic line has been kept for research purposes. Within this line, Vitamin C deficiency presented classical signs of “Scurvy” in sows and the skeletal system of their off-spring in as little as 25-30 d after removal of ascorbic acid from the diet (Wegger and Palludan, 1994). A re-view by Mahan et al. (2004) indicated there may be key time points during the reproductive cycle that supple-mentation C would be beneficial, but the relative body of work indicates no additional benefits of supplement-ing vitamin C to sows that are typical. Because vitamin C has not been recognized to be important for supplementation in typical swine, very little swine research has occurred for this vitamin. Has vitamin C metabolism unknowingly been altered in modern, swine genetic improvement programs? Or, are some prolific sows unable to produce adequate amounts for their optimum health and the health of their prog-eny? As previously discussed, zearalenone contamina-tion can add additional weight and length to the repro-ductive tract, but it also causes oxidative stress in the

liver. Shi et al. (2017) demonstrated that supplementing gilts with 150 mg/kg of Vitamin C was able to alleviate the oxidative stress damage of zearalenone contaminat-ed feed. Thus, their research confirms a potential benefit with supplemental Vitamin C in swine diets, particular-ly when presented with a mycotoxin contamination. Vitamin C is also a key regulator in collagen forma-tion. The lack of vitamin C can lead to impaired collagen formation or, at a minimum, reduced strength of colla-gen formed structures, such as ligaments. As described before, improper collagen and elastin metabolism have been found to be related to pelvic floor disorders and prolapses (POP) in women. Findik et al. (2016) demon-strated that Vitamin C supplementation during preg-nancy in rats may have aided in the prevention of POP by increasing the production of type I and type III col-lagen in the cardinal and uterosacral ligaments.

Vitamin D Although a requirement for dietary vitamin D sup-plementation is recognized for domestic swine, much of the previous research used to establish published re-quirement estimates has been based on a rather limited number of studies. Only recently have there been stud-ies evaluating the effects of differing levels and sources of vitamin D supplementation on reproductive and progeny performance, immune function, the incidence of OC, or other variables related to greater health, lon-gevity, or reproductive efficiency. Lauridsen et al. (2010) reported that the number of stillborn pigs per litter was reduced when maternal diets were supplemented with 1,400 or 2,000 IU vitamin D/kg rather than 200 or 800 IU/kg. When an increased level of vitamin D supple-mentation was provided to gilts throughout gestation and lactation using the addition of 2,000 IU of 25(OH)D3/kg to a diet already containing 2,000 IU vitamin D3/kg, Zhou et al. (2016) reported that the higher level of supplementation (using the combination of vitamin D forms) improved the reproductive performance, milk quality and bone status of sows, as well as the bone char-acteristics of the newborn piglets. Using 2 dietary vita-min D treatments similar to that of Zhou et al. (2016) (i.e., basal level of vitamin D3 vs. basal diet + 2,000 IU 25(OH)D3/kg), Sugiyama et al. (2013) reported that the growth performance and bone density was not differ-ent from 6 to 110 kg BW, but serum levels of 25(OH)D3 were increased, and the incidence of osteochon-drotic lesions of the humerus and femur at 110 kg BW was greatly reduced, for the pigs fed added vitamin D in the form of 25(OH)D3. Higher levels of vitamin D than those recommended for preventing classical deficien-cies are needed to optimize health and performance,

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but the 25(OH)D3 form of the vitamin is very effective for increasing the level of vitamin D supplementation.

Trace Minerals - Organic vs Inorganic Sources It has been well established that there is a wide range of trace mineral sources with varying bioavailability for animals. Chelated (organic) minerals have been inten-sively studied in both cows and sows for their capabil-ity to reduce lameness and claw lesions. The potential causes of claw lesions associated with lameness are ge-netics, flooring, nutrition, injury and other factors. As a sow’s BW increases, claw sole area increases quadrati-cally while claw volume increases linearly. Also, the rear inner claws generally remain smaller in sole area and volume than the other claws (Figure 3, Bradley, 2010). As reported by Bradley (2010), organic trace mineral supplementation did not impact overall quantity of le-sions presence, but did appear to reduce the severity of claw hemorrhages found, which were most likely due to injury. Further work has demonstrated organic trace mineral supplementation to lower the frequency and/or severity of lesions in sows (Lisgara et al., 2016).

Boron Boron is the fifth element on the periodic table and it belongs to group 13, that also contains aluminum. Most ICP analysis can detect aluminum, but are not programmed to analyze boron. Similarly, there is no approved boron food additive for feed. It was accepted

that boron was essential for plant nutrition as early as 1926 (Sommer and Lipman), which was considered unique until the 1980’s because to date it did not ap-pear essential for livestock. In a chick study, Hunt and Nielsen (1981) suggested that it stimulated growth and partially prevented leg abnormalities in Vitamin D de-ficient chicks. More recently using swine, the supple-mentation of boron at 25 ppm in a grower diet reduced the incidence of lameness (Johnson et al., 2005).

Macronutrient Considerations—Collagen Formation Protein and/or amino acids and energy require-ments, in general, have been studied in much greater depth than micronutrients for sows. However, NRC Swine (2012) cited only 4 trials to determine the SID lysine values for gestating sows, and 3 of those studies were published 40+ years ago. There were 10 studies for lactating sows, but the most recent publication was from 2001. Swine diets are normally only formulated to the fourth or fifth limiting amino acid (lysine, tryptophan, threonine, valine, and isoleucine), while nonessential amino acids are rarely considered. For instance, glycine is not considered a limiting amino acid, but is important for collagen formation. Glycine comprises one-third of the amino acid residues of collagen. In metabolism, gly-cine is utilized in two ways: 1) building block for por-phyrins, purine bases, creatine, glutathione, bile salts, and hippuric acid; and 2) for amino acid synthesis of

Figure 3. A: Diagram of different claw measurements. B: Regression plot of how a sow’s sole area (SA) changes with BW (SA = Claw Width x Sole Length. C: Regression plot of how a sow’s claw volume (CV) changes with BW (CV = SA x heel height). L = left foot, R = right foot, FI = Front Inner Claw, FO = front outer claw, RI = rear inner claw, RO = rear outer claw. (Bradley, 2010).

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proteins, especially collagen. Within human nutrition, Meléndez-Hevia et al. (2009) reported that glycine is a semi-essential amino acid and should be supplemented at higher levels to promote healthy metabolism. The au-thors detailed the inability for glycine to be adequately present for metabolic needs, but especially for proper collagen synthesis, which may be further compromised during pregnancy and in aging populations. Thus, as we consider lower crude protein diets in growing animals and sows with and the use of synthetic essential amino acids, the non-essential amino acids should be evaluat-ed beyond traditional growth performance and repro-ductive parameters.

Conclusions Within this review of pelvic floor disorders and prolapses in humans, there are numerous possibilities to influence the condition of supportive ligaments via nutrition. Furthermore, stressors within a sow’s lifetime may have a negative effect on her longevity. A “perfect storm” could be occurring within today’s sow herds to create the increases in sow mortality and uterine pro-lapses. Knowledge and research to solve the current is-sues of concern are mostly lacking, and the traditional methods of measuring sow reproductive traits for great-er profitability are insufficient to solve the existing and future problems. This review has provided some unique insights into opportunities for improving our under-standing of nutritional requirements that may be help-ful in reducing the risks for sow mortality.

ReferencesBradley, C. L. 2010. The efficacy of organic trace mineral

supplementation to developing gilts and reproduc-tive sows on reproduction efficiency, lameness, and longevity. (PhD dissertation) University of Arkan-sas, Fayetteville.

Day, C. (2017). Increase in U.S. sow mortality a real mys-tery. National Hog Farmer (May issue). http://www.nationalhogfarmer.com/animal-health/increase-us-sow-mortality-real-mystery.

Findik, R. B., F. Ilkaya, S. Guresci, H. Guzel, S. Karabulut, and J. Karakaya. 2016. Effect of vitamin C on colla-gen structure of cardinal and uterosacal ligaments during pregnancy. Euro. J. Obstetrics Gynecology Repro. Bio. 201: 31-35.

Hahn, L., L. Wang, Q. Wang, H. Li, and H. Zang. 2014. Association between pelvic floor organ prolapse and stress urinary incontinence with collagen. Exp. Therapeutic Med. 7: 1337-1341.

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Meléndez-Hevia, E., P. De Paz-Lugo, A. Cornish-Bowden, and M. L. Cardenas. 2009. A weak link in metabolism: the metabolic capacity for glycine bio-synthesis does not satisfy the need for collagen syn-thesis. J. Biosci. 34(6): 853-872.

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Reese, M. E., and E. Casey. 2015. Chapter 2: Hormonal Influence on the Neuromusculoskeletal System in Pregnancy. In C. M. Fitzgerald and N.A. Segal (Eds.), Musculoskeletal Health in Pregnancy (pp 19-39). Switzerland: Springer International.

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Shi, B., Y. Su, S. Chang, Y. Sun, X. Meng, and A. Shan. 2017. Vitamin C protects the piglet liver against zearalenone-induced oxidative stress by modulat-ing expression of nuclear receptors PXR and CAR and their target genes. Food Funct. DOI: 10.1039/C7FO01301A.

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Zhao, X. J., X. Y. Wang, J. H. Wang, Z. Y. Wang, L. Wang, and Z. H. Wang. 2015. Oxidative stress and imbal-ance of mineral metabolism contribute to lameness in dairy cows. Biol. Trace Elem. Res. 164: 43-49.

Zhou, H., Y. Chen, Y. Zhuo, G. Lv, Y. Lin, B. Feng; Z. Fang, L. Che, J. Li, S. Xu, and D. Wu. 2017. Effects of 25-hydroxycholecalciferol supplementation in ma-ternal diets on milk quality and serum bone status markers of sows and bone quality of piglets. Anim. Sci. J. 88(3):476-483.

Zong, W., S. E. Stein, B. Starcher, L. A. Meyn, and P. A. Moalli. 2010. Alteration of vaginal elastin metabo-lism in women with pelvic floor organ prolapse. Obestet. Gynecol. 115(5): 953-961.

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Impact of Dietary Amino Acid Balance on Nitrogen and Energy Efficiency in Sows:

from Theory to PracticeTrottier, N. L. and S. Zhang

Department of Animal Science Michigan State University

East Lansing, MI 48824 Phone: 517-432-5140

[email protected]

J. S. Johnson USDA-ARS Livestock Behavior Research Unit

Purdue University West Lafayette, IN 47907

SummaryAggressive reduction in dietary crude protein and crystalline amino acid supplementation to im-prove dietary amino acid balance minimizes urea-nitrogen synthesis and ammonia emission with-out compromising lactation performance. Improving dietary amino acid balance also increases milk yield and true milk protein yield in peak lactation, and amino acid and global nitrogen utili-zation efficiency for milk protein production. Individual amino acid efficiencies are dynamic, and correct use of these efficiency values is critical for accurate prediction of amino acid requirements. Future dietary formulations using reduced crude protein diets to minimize nitrogen excretion and ammonia emission will require amino acid requirements based on near maximum biological ef-ficiency estimates.

Introduction The use of crystalline amino acids (CAA) in con-junction with reduction in dietary crude protein (CP) allows for improvement of dietary amino acid (AA) balance and increases whole body nitrogen (N) and en-ergy utilization efficiency for milk production in sows (Huber et al., 2015; Zhang et al., 2018). Improvement in dietary AA balance reduces N excretion and am-monia emission (Hubert et al., 2015; Chamberlin et al., 2015b). The global improvement in N utilization is due to incremental change in utilization efficiency of indi-vidual dietary AA (Huber et al., 2015). Except for that of lysine (Lys), utilization efficiencies of the nine remain-ing essential AA have not been systematically estimated (NRC, 2012). Prediction of dietary AA requirement through the use of models (NRC, 2012) are dependent on valid AA efficiency estimates. Amino acid efficien-cies however are dynamic, (White et al., 2016), and de-pendent on their own dietary concentrations and that of total N. This plasticity complicates the application of AA efficiency for prediction of requirements. The ob-jectives of this presentation are to provide a summary

of the data we have thus far collected from studies with sows fed diets with improved AA balance. We will dis-cuss the impact of such diets on sow performance, urea excretion and ammonia emissions, AA efficiencies and future direction on implementation strategies of AA ef-ficiency estimates for prediction of AA requirements.

Sow Performance and Milk Protein Across studies, results show that feeding reduced CP diets containing as low as 12.56% CP with CAA to meet the SID requirement of the limiting AA does not impact piglet ADG, litter growth rate, and sow feed intake (Table 1). In the studies by Chamberlin et al. (2015b), Huber et al. (2015) and Zhang et al. (2018), sows lost more weight and retained less N in their body when fed the lowest level of CP. The greater loss in sow body weight (BW) appears to be associated with greater body protein loss rather than body fat. It is possible that other AA may have been limiting, leading to body pro-tein mobilization, and reflecting an underestimation of dietary requirements in NRC (2012) for total N or an essential AA for which the dietary concentrations were

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Table 1. Performance of lactating sows fed diets reduced in crude protein (CP) concentration with supplemental crystalline amino acids.1

StudyCP, %

SID Lys, %

SID Thr, %

SIDM+C,

%SID Trp,

%

Litter gain,

kg

Piglet ADG, g/d

Sow BW loss, g/d

Sow fat change

∆

Sow protein

∆Manjarin et al., 2012 17.52 1.11 0.69 0.55 0.21 1.71 214 228 - -

13.53 0.85 0.53 0.42 0.16 2.26 282 232 - -Huber et al., 2015 17.62 0.74 0.59 0.50 0.18 1.86 186 414 - -

14.63 0.74 0.59 0.50 0.18 2.18 221 433 - -Huber et al., 2015 16.03 0.74 0.59 0.50 0.18 2.32 238 143 −0.1 +0.2

15.70 0.74 0.59 0.50 0.18 2.53 256 176 −0.2 −0.814.29 0.74 0.59 0.50 0.18 2.41 243 190 −0.1 −1.213.22 0.74 0.59 0.50 0.18 2.60 260 285 −0.2 −2.7

Chamberlin et al., 2015a 17.16 0.78 0.53 0.48 0.18 2.53 262 270 - -14.79 0.78 0.49 0.42 0.15 2.64 278 413 - -12.56 0.78 0.49 0.41 0.15 2.56 258 358 - -

Chamberlin et al., 2015b 17.162 0.78 0.53 0.48 0.18 2.60 265 500 −0.06 -12.562 0.78 0.49 0.41 0.15 2.80 279 300 −0.13 -17.163 0.78 0.53 0.48 0.18 2.40 244 700 −0.15 -12.563 0.78 0.49 0.41 0.15 2.30 238 800 −0.10 -

Zhang et al., 2018 18.74 0.90 0.61 0.54 0.21 2.43 251 62 −0.054 +5.46 13.78 0.90 0.58 0.49 0.17 2.56 255 395 −0.175 −11.26

1 NE=2,580 to 2,600 kcal/kg.2 Sows were housed under thermal neutral environmental temperature.3 Sows were housed under thermal heat stress environmental temperature.4 Corresponds to 77 g/d body fat loss.5 Corresponds to 312 g/d body fat loss.6 Body protein loss, g/d.

lowest in the lowest CP diets (e.g., arginine (Arg), histi-dine (His), leucine (Leu), phenylalanine (Phe), and Phe + tyrosine (Tyr). Depending on the stage of lactation, milk casein concentration and casein to true protein (TP) ratio are either unaffected by AA balance, or greater. Both Huber et al. (2015) and Chamberlin et al. (2015b) reported 25% increases in casein concentration and yield (data not shown), and casein:TP in peak lactation (Table 2).

Overall Efficiency of Nitrogen Utilization Utilization of dietary N for milk production increas-es in response to decreasing dietary CP concentration and increasing CAA supplementation, provided that the supply of potentially limiting AA meet their estimat-ed requirements. In both studies by Huber et al. (2015) and Chamberlin et al. (2015a and 2015b) feeding re-duced CP diets lowered milk urea-N over 2-fold in early lactation and by more than 5-fold in peak lactation (Fig-ures 1 and 2). In both studies, milk urea-N concentra-tion from early to peak lactation did not change in sows fed the reduced CP diet, but nearly doubled in sows fed a non-reduced CP diet. Similarly, in early lactation, plas-ma urea-N of sows fed a low CP diet was nearly half that of control, and in peak lactation, plasma urea-N of sows fed a low CP diet was reduced, indicating greater uti-lization of N compared to the low-CP fed sows. These changes are equally reflected in the urinary N excre-

tion response (Figure 2, right panel) and a remarkable decrease in ammonia emissions under either thermal neutral or thermal heat stress environments (Figure 3). The urea cycle is an energy expensive process, whereby two molecules of ammonia, and one mole-cule of carbon dioxide are used for the synthesis of one molecule of urea ((NH2)2CO). The reduction in MUN coupled with the reduction in plasma urea-N without a decrease in true protein or casein yield suggests sows fed near optimum AA balance are metabolically more efficient and that energy cost associated with excess AA catabolism and urea synthesis can be minimized. The recent study by Zhang et al. (2018) showed that feeding sows with a CAA-supplemented diet containing 13.78% CP compared to a 18.74% CP diet increased milk N pro-duction per unit of ME and NE in early (20% increase) and peak (over 30% increase) lactation periods, with a drastic increase in efficiency of dietary N into milk N (Table 3).

Efficiency of Amino Acid Utilization Earlier, Manjarín et al. (2012) reported that decreas-ing dietary CP by 4% (from 17.5% to 13.5%) and meeting the limiting AA requirement via CAA supplementation increased the efficiency of Lys and Arg utilization by the mammary gland. As discussed previously, aggressive reduction in dietary CP and CAA inclusion rates can minimize serum and milk urea-N and ammonia emis-

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Table 2. Milk casein concentration and milk casein to true protein ratio from lactating sows fed diets reduced in crude protein concentration with supplemental crystalline amino acids.

Study CP, %Early lactation, d 3-7 Peak lactation, d 14-18

Casein, % Casein:TP1 Casein, % Casein:TPHubert et al., 2015 16.03 3.63 0.70 3.29 0.79

15.70 3.30 0.65 3.21 0.7714.29 3.64 0.68 3.38 0.8413.22 3.50 0.67 3.41 0.77

Chamberlin et al., 2015a 17.16 3.70 0.78 2.85 0.6714.79 3.25 0.66 3.15 0.8012.56 3.76 0.79 3.81 0.90

Chamberlin et al., 2015b 17.161 2.77 0.69 2.65 0.7612.561 2.81 0.74 2.45 0.7317.162 2.65 0.76 2.57 0.7312.562 2.45 0.73 2.64 0.75

Zhang et al., 2018 18.74 4.522 - 4.462 -13.78 4.232 - 4.522 -

1 True protein.2 True protein concentration, %.

Figure 2. Milk (left panel) and urinary (right panel) urea nitrogen (mg/dL and g/d, respectively) from sows exposed to heat stress (HS) and thermo-neutral temperature (TN) and fed a Low protein diet or a Control diet during lactation. From Chamberlin et al. (2015b).

Figure 1. Milk urea concentration (mg/dL) in sows fed diets containing 17.55, 15.25 and 12.98 % CP (Control, Med and Low, respectively), in early (d 3-7) and peak (d 14-17) lactation. From Chamberlin et al. (2015a).

sion without compromis-ing lactation performance, indicating that the dietary AA balance is improved. Within this range of di-etary CP concentrations, improving dietary AA bal-ance also led to increased estimated milk yield and estimated true milk protein yield during peak lactation, and improved N utilization efficiency for milk protein production. The global im-provement in N utiliza-tion is due to incremental change in utilization ef-ficiency of individual dietary AA (Huber et al., 2015). Amino acid efficiencies are dynamic (White et al., 2016) as depicted in Table 4. Amino acid efficiency estimates across dietary CP concentrations and CAA inclusion rates generally increase, and quite considerably for some AA (Arg, His, Ile, Leu) with dietary improvement in AA balance. Note that efficiency estimates for Val re-ported by Huber et al. (2015) decrease with reduction in CP while those of Zhang et al. (2018) increase. This discrepancy stems from the fact that the highest CP diet in Zhang et al. (2018) did not contain crystalline valine (Val) while that of Huber et al. (2015) contained crystal-line Val and was likely over Val requirement. The NRC (2012) efficiency estimates were not systemically de-termined, except for that of Lys, and therefore it is not

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surprising that many AA differ quite substantially from those reported by Huber et al. (2015) and Zhang et al. (2018). In addition, AA efficiency values from Zhang et al. (2018) are notably higher than those of Huber et al. (2015) for peak lactation, and this may be related to the use of sow BW loss over a 21-d period rather than the specific peak lactation period of d 14 to 17. Con-sequently, Trp for instance is over 100%. Hence, for practical purpose in estimating requirement using a modeling approach, efficiency esti-mates generated over a 21-d period would be more relevant and biologi-cally correct. Maximum biological efficiency estimates are needed to predict AA requirement. Efficiency estimates below the maximum bio-logical efficiency leads to over esti-mation of AA requirement. We have conducted a meta-analysis using the current available studies on AA requirements for lac-tating sows and generated AA effi-ciency estimates. Our estimated val-ues were too low to use in practical formulation, except for that of Lys, which was in line with the estimated value of Huber et al. (2015), Zhang et al. (2018) and NRC (2012). This makes sense because the great ma-jority of studies are designed to de-termine the minimum Lys require-ment. This exemplifies the need to conduct systematic experiments with formulations targeted to gener-ate maximum biological efficiency values for Thr, methionine+cystien (Met + Cys), Trp, Ile, Phe, Val, and Leu. Accurate efficiency values for Arg and

Table 4. Efficiencies of SID amino acid utilization for milk production in sows fed diets with reduced crude protein concentrations and supplemental crystalline amino acids, %.

ItemCrude protein concentration, %

NRC319.241 16.582 15.352 14.152 13.781 12.892

Early lactation (d 3-7)Arg 30.1 35.4 36.8 40.2 50.6 39.5His 52.2 58.4 65.7 68.6 69.8 73.6Ile 43.5 48.9 53.0 51.3 62.5 50.6Leu 43.2 46.4 53.9 53.7 64.2 59.6Lys 60.1 67.7 71.0 64.3 61.1 62.6Met 64.9 55.7 60.1 48.4 57.6 44.2Met + Cys 53.1 59.2 66.0 58.9 60.5 58.4Phe 37.4 42.0 48.2 49.6 48.7 53.1Phe + Tyr 44.3 55.3 60.4 61.2 61.6 60.4Thr 55.7 58.6 64.0 56.7 60.0 54.1Trp 62.0 49.9 54.3 48.7 87.8 51.8Val 51.3 44.5 46.6 42.6 50.7 39.4N 44.4 55.3 60.4 61.2 62.7 60.4Essential AA 52.2 - - - 62.5 -

Peak lactation (d 14-18)Arg 33.5 31.7 41.0 43.8 69.9 53.6 81.6His 58.1 51.8 64.8 67.8 93.8 81.5 72.2Ile 48.4 43.1 50.6 51.0 83.6 54.3 69.8Leu 48.2 41.0 49.9 50.8 85.6 59.5 72.3Lys 67.0 59.9 68.4 65.4 82.1 67.6 67.0Met 71.8 50.0 58.9 48.8 77.1 47.0 67.5Met + Cys 59.0 52.6 61.6 57.1 80.7 57.9 66.2Phe 41.7 37.0 45.8 48.5 65.2 56.9 73.3Phe + Tyr 49.3 48.9 59.6 60.3 82.2 68.2 70.5Thr 61.9 52.6 60.0 57.4 80.2 59.3 76.4Trp4 68.4 44.2 48.8 46.6 114.0 52.3 67.4Val 57.1 39.3 45.0 44.5 68.0 45.0 58.3N 49.8 48.9 59.6 60.3 84.7 68.2 75.9Essential AA 58.1 - - - 83.3 - -

1 Zhang et al., 2018. Unpublished.2 Huber et al., 2015.3 NRC, 2012. Values are over a 21-d lactation period and are not restricted to the peak

lactation period.

Table 3. Energetic utilization of milk nitrogen deposition.

Days in lactation CP

Milk N/ME intake,

mg/kcal1

Milk N/NE intake,

mg/kcal2

N output/ N intake,

%3

4 to 7 18.74 3.62 4.83 40.413.78 4.45 5.86 67.5

14 to 17 18.74 3.74 3.73 41.713.78 4.98 4.89 56.5

Zhang et al. (2018).1 SEM = 0.252 SEM = 0.353 SEM = 3.8

Figure 3. Air ammonia production in individual lactating sows and their litters. Sows were fed diets containing 17.55 and 12.98% CP, and housed under either thermal neutral (TN) or thermal heat stress (HS) environments. From Chamberlin et al. (2015b).

His remain debatable because of de novo synthesis and de novo availability, respectively.

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Conclusion Aggressive reduction in dietary CP and CAA sup-plementation to improve dietary AA balance mini-mizes urea-N synthesis and ammonia emission with-out compromising lactation performance. Improving dietary AA balance also increases milk yield and true milk protein yield in peak lactation, and AA and global N utilization efficiency for milk protein production. In-dividual AA efficiencies are dynamic, and correct use of these efficiency values is critical for accurate prediction of AA requirements. Future dietary formulations using reduced CP diets to minimize N excretion and ammo-nia emission will require AA requirements based on near maximum biological efficiency estimates.

ReferencesChamberlin, D.P., D.W. Rozeboom, S. Erwin, and N.L.

Trottier. 2015a. Lactation performance in sows fed diets with graded levels of crystalline amino acids as substitute for crude protein at lysine requirement. Journal of Animal Science 93 (E-Suppl. 2):21.

Chamberlin D.P., W.J. Powers, D.W. Rozeboom, T.M. Brown-Brandl, S. Erwin, C. Walker, and N.L. Trot-tier. 2015b. Impact of reduced dietary crude protein concentration with crystalline amino acid supple-mentation on lactation performance and ammonia emission of sows housed under thermo neutral and thermal heat stress environments. Journal of Animal Science. 93 (E-Suppl. 2):19.

Huber, L., C.F.M. de Lange, U. Krogh, D. Chamberlin, and N.L. Trottier. 2015. Impact of feeding reduced crude protein diets to lactating sows on nitrogen uti-lization. Journal of Animal Science. 93:5254–5264.

Manjarín, R., V. Zamora, G. Wu, J.P. Steibel, R.N. Kirk-wood, N.P. Taylor, E. Wils-Plotz, K. Trifilo, and N.L. Trottier. 2012. Effect of amino acids supply in reduced crude protein diets on performance, ef-ficiency of mammary uptake, and transporter gene expression in lactating sows. Journal of Animal Sci-ence. 90:3088-3100.

NRC (National Research Council). 2012. Nutrient Re-quirements of Swine. Eleventh revised edition. Na-tional Academy Press, Washington, DC.

White, R. R., S. Zhang, N. Regmi, and N. L. Trottier. 2016. Quantifying variable amino acid efficiencies in lactating sows. Journal of Animal Science. 94 (E-Suppl. 2):17.

Zhang, S., J. Johnson, M. Qiao, J. Liesman, and N. L. Trottier. 2018. Feeding a diet with a near optimal amino acid profile improves energy utilization for milk production in lactating sows. Journal of Animal Science. 96 (E-Suppl. 2):87.

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Copper Levels and Sources for SowsNing Lu and Merlin Lindemann

Department of Animal and Food Sciences University of Kentucky, Lexington 40546


SummaryIn the present study, 31 crossbred gilts were fed experimental diets to determine the effects of dietary Cu sources [copper sulfate (CuSO4) or tribasic copper chloride (TBCC)] and levels (20, 120, or 220 mg/kg) on performance and health of sows and their progeny. Sows fed TBCC diets had greater ad-justed weaning weight and adjusted lactation weight gain for the litter and/or the individual piglet (P < 0.10), respectively, when compared to sows that received CuSO4 diets. Increasing dietary Cu level linearly increased (P = 0.06) live born piglet weight. Sows fed TBCC diets had greater apparent total tract digestibility of dry matter and nitrogen during lactation (P < 0.05). Increasing Cu levels increased levels of milk fat and Cu (linear, P < 0.05); but linearly decreased lactose and Zn levels (P < 0.05). Lactating sows fed TBCC diets had a greater activity of Cu/Zn superoxide dismutase (SOD) and ceruloplasmin in serum than those fed CuSO4 diets (P < 0.05). Increasing dietary Cu levels in-creased total and Cu/Zn SOD activity for lactating sows (linear, P < 0.05). Sows fed TBCC diets had lower concentrations of Cu (P = 0.04), but greater concentrations of iron and manganese (P < 0.05) in the liver, when compared to those fed CuSO4 diets. In addition, liver Cu concentration increased with increasing dietary Cu levels (linear and quadratic, P < 0.05). Increasing sow dietary Cu levels did not influence tissue trace mineral concentration in neonatal piglets, but resulted in the elevation of Cu concentrations in the liver of weanling piglets (linear, P < 0.0001). Results of this study suggest that the source of Cu affects reproductive performance, and that higher dietary Cu levels in sow diets than the NRC (2012) requirement estimates are beneficial to sows and piglets without showing any apparent adverse effects.

Introduction Sow productivity is a key determinant of swine enterprise profitability. Sow productivity in the major pork producing countries, including China, the Euro-pean Union, and the U.S., has increased by 11 to 28% from 2001 to 2013, regarding weaned or finished pigs produced per sow per year (Lu, 2018). However, greater sow productivity inevitably leads to increased mobili-zation of minerals from body tissues, and reproductive capacity can be compromised if the mineral needs for reproductive demands exceed body stores and dietary intake (Mahan, 1990). It has been demonstrated that body mineral contents, including calcium, phosphorus, magnesium, potassium, sodium, aluminum, zinc, and copper declined in sows that had completed three pari-ties as compared to those of similarly aged, nongravid gilts (Mahan and Newton, 1995). Copper is required by pigs to serve many biological roles in the body, such as supporting iron metabolism,

protecting tissues from oxidative damage, and main-taining immunity (Hill and Spears, 2001). The latest edi-tion of the NRC estimates that the Cu requirement of growing pigs is 3 to 6 mg/kg and for gestating and lactat-ing sows is 10 and 20 mg/kg, respectively (NRC, 2012). Moreover, pharmacological concentrations of Cu (125 to 250 mg/kg) have been extensively studied and dem-onstrated to enhance growth performance of weanling and growing pigs (Cromwell, 2001). Regarding breed-ing herds, dietary supplementation of pharmacological levels of Cu (250 mg/kg) in gestation and lactation diets from Parity 1 to 6 has been demonstrated to improve piglet weight at birth and weaning, as well as increase Cu concentrations in sow liver and kidney (Cromwell et al., 1993). However, no study has been reported to as-sess the effects of high dietary Cu from different sources on reproducing sows. Therefore, the objective of the present study was to determine the long-term effects of both dietary Cu source and level on performance and health of sows and their progeny.

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Experimental ProceduresAnimals and experimental design A total of 31 eight-month-old gilts [Yorkshire; (York-shire × Landrace) × Duroc] were provided with a com-mon gestation diet containing 8 mg/kg Cu as copper sulfate (CuSO4) from breeding until their pregnancies were confirmed at 55 ± 2 d post-breeding. Upon con-firmation of pregnancy, gilts were allotted to a 2 × 3 fac-torial arrangement in a completely randomized design with 2 Cu sources [CuSO4 or tribasic copper chloride (TBCC)] and 3 Cu levels (20, 120, and 220 mg/kg). All experimental animals received their respective diets until they were removed from the study or until they weaned their 4th litter.

Experimental Diets Corn-soybean meal diets were formulated to meet or exceed NRC (2012) nutrient requirement estimates for gestating and lactating sows. Diets 1 to 3 were de-signed to contain 20, 120, and 220 mg/kg of Cu as TBCC, respectively; Diets 4 to 6 were designed to contain 20, 120, and 220 mg/kg of Cu as CuSO4 , respectively. Tita-nium dioxide was added at a level of 0.30% by replacing an equal amount of corn as an indigestible marker.

Data and Sample Collection The number and weight of total piglets born, born alive, and stillborn were recorded within 12 h after far-rowing; and the number and weight of piglets at wean-ing was also recorded. The weight of the piglet and litter at weaning were adjusted to a common 21-d lactation as described by Jang et al. (2017). Diet samples were collected at the feed mill for every mixing batch. Fecal samples were collected for lactating sows during the 2nd and 3rd parity from d 15 to 17 post-farrowing by grab sampling. Blood samples were collected via vena cava puncture from sows during the 2nd to 4th parity at d 15 post-farrowing. During the 3rd and 4th parity, milk samples were collected at d 15 post-farrowing from each sow after intra-venous injection of 1 mL oxytocin. At the beginning of the study, 6 open gilts from the same breeding group were slaughtered for tissue collection (liver, heart, right kidney, and both ovaries). After completing at least 3 parities, sows were slaughtered for the same tissue collection. One piglet was sacrificed at birth, and another one at weaning for tissue collection (liver, kidneys, and heart) for the 3rd and 4th litter of each sow.

Table 1. Distribution of number of litters1, 2

Copper source: Tribasic copper chloride

Copper sulfateCopper level, mg/kg: 20 120 220 20 120 220

Item Diet No.: 1 2 3 4 5 6No. of gilts allotted 5 6 5 5 5 5No. of litters

1st-parity litters 4 6 5 5 5 52nd-parity litters 4 6 5 5 5 53rd-parity litters 3 6 5 4 4 44th-parity litters 3 4 4 4 2 4Total 14 22 19 18 16 18

Average no. of litters/sow 3.5 3.7 3.8 3.6 3.2 3.61 No significant difference across treatments.2 One gilt from Diet 1 did not farrow throughout the experiment, and one sow from

Diet 3 died after completion of the 3rd parity.

Sample Analysis Feed and fecal samples were analyzed for dry matter (DM), gross energy (GE), ether extract (EE), and nitro-gen. Malondialdehyde (MDA) concentration, superox-ide dismutase (SOD) activity, and ceruloplasmin (Cp) activity were analyzed in serum samples. Milk composi-tion analysis was conducted using Foss Milkoscan™ FT 120 device.

Statistical Analysis All data were subjected to ANOVA using the GLM procedure in SAS (Statistical Analysis System, Cary, NC, USA) for a completely randomized design. The individual litter served as the experimental unit and results are reported as least squares means. The statis-tical model included the Cu source, Cu level, and sow parity, as well as their interactions. With the purpose of achieving appropriate statistical power, the α level used for determination of statistical significance was set at 0.10 for sow and litter performance data (α level at 0.15 for tendency), and set at 0.05 for all other data (α level at 0.10 for tendency).

ResultsReproductive Performance A total of 107 litters were farrowed in the present study (Table 1). Chi-square analysis showed that the lit-ter distribution did not differ across dietary treatments. One sow from Diet 1 was culled because of reproduc-tive failure (no litter farrowed), and another sow from Diet 3 was removed because of death from gastric tor-sion (after completion of 3 parities). Sows fed the TBCC diets had more stillborn piglets (P = 0.03), greater adjusted weaning weight of piglets (P = 0.07), and adjusted litter (P = 0.10) and piglet (P = 0.05) weight gain than CuSO4 fed sows (Table 2). In addition, tendencies for greater litter weight of total born piglets (P = 0.11), piglet weight at weaning (P = 0.13), piglet weight

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gain during lactation (P = 0.12), and adjusted litter weight at weaning (P = 0.14) were observed for TBCC fed sows compared to CuSO4 fed sows. Litter performance (litter size, litter weight, and piglet weight) was not affected by Cu level except for the weight of total piglets born (lin-ear. P = 0.12) or born alive (linear, P = 0.06). An inter-action of Cu source and level was detected on adjusted piglet weight at weaning (P = 0.06), which showed a lin-ear increase with increasing Cu levels in TBCC diets (P = 0.02) but no response to Cu level within CuSO4 diets (P > 0.28). Moreover, the interaction was also observed on adjusted piglet weight gain (P = 0.02), which showed an increasing linear response to increasing copper level (P = 0.02) within TBCC diets but a decreasing linear re-sponse within CuSO4 diets (P = 0.05).

Apparent Total Tract Digestibility Sows fed TBCC diets had greater apparent total tract digestibility (ATTD) of DM, nitrogen (P < 0.05), and GE (P < 0.10) when compared to the ones fed CuSO4 diets (Table 3). The increasing dietary Cu levels elevated the ATTD of DM (linear, P < 0.05) and GE (linear, P < 0.06).

Hematology and Antioxidant Status Lactating sows fed TBCC diets had greater activity of total SOD (P = 0.08), Cu/Zn SOD (P = 0.04), and Cp (P = 0.01) compared to sows fed the CuSO4 diets (Table 4). The activity of total and Cu/Zn SOD increased linearly (P < 0.03) with increasing dietary Cu levels. In addition, the activity of Mn SOD and Cp quadratically changed with increasing dietary Cu levels (P < 0.05). However, hematocrit, hemoglobin, or MDA were not affected by dietary Cu source or level (P > 0.23).

Milk Composition Milk from sows that were fed TBCC diets had great-er levels of protein than that from sows that were fed CuSO4 diets (P = 0.02, Table 5). Furthermore, the TBCC fed sows tended to have greater levels of fat, GE, and to-tal solids, but a lower level of lactose in milk than the CuSO4 fed sows (P < 0.10). With increasing dietary Cu levels, concentrations of fat and Cu (P < 0.05), as well as gross energy and total solids (P < 0.10) linearly increased in milk. However, linearly decreased levels of lactose (P

Table 2. Effects of dietary copper source and level on litter performance1


Copper sulfate

SEMP valuesCopper level, mg/kg: 20 120 220 20 120 220

Item Diet No.: 1 2 3 4 5 6 Source LevelNo. of litters 8 15 12 13 11 8Lactation length, d 21.1 20.3 19.8 20.7 20.6 20.0 0.5 0.96 0.20Litter size

Total born 10.67 10.64 11.51 10.93 10.02 9.83 0.83 0.33 0.83Live born 10.00 9.77 10.36 10.47 9.77 9.61 0.83 0.89 0.85Stillborn 0.56 0.81 1.16 0.47 0.33 0.22 0.26 0.03 0.81Weaning 9.11 8.58 9.09 9.17 8.45 8.28 0.70 0.61 0.66

Litter weight, kgTotal born 17.12 15.96 18.52 14.86 15.57 16.34 1.20 0.11 0.35Live born 16.26 14.84 16.77 14.32 15.25 15.54 1.25 0.37 0.66Weaning 58.38 55.30 58.68 57.02 53.02 47.66 4.52 0.19 0.59Litter gain6 43.94 42.01 44.10 44.20 39.45 34.25 3.54 0.17 0.40

Piglet weight, kgTotal born5 1.53 1.54 1.67 1.41 1.56 1.59 0.10 0.46 0.30Live born2 1.54 1.56 1.74 1.42 1.57 1.61 0.10 0.31 0.17Weaning 6.40 6.48 7.10 6.35 6.41 5.88 0.35 0.13 0.95Piglet gain4, 6 4.82 4.92 5.33 4.91 4.80 4.24 0.29 0.12 0.95

Adjusted weight, kgLitter weight at weaning 57.80 56.59 61.14 57.52 53.68 49.01 4.17 0.14 0.79Piglet weight at weaning3 6.34 6.65 7.41 6.44 6.49 6.06 0.31 0.07 0.57Litter weight gain6 43.36 43.31 46.56 44.70 40.11 35.60 3.14 0.10 0.63Piglet weight gain4, 6 4.76 5.08 5.64 5.00 4.88 4.41 0.24 0.05 0.84

1 Data were collected from the 2nd to 4th parity litters.2 Linear response to copper level (P = 0.06).3 Copper source × copper level interaction (P = 0.06). A linear response to increasing copper level (P = 0.02) within tribasic

copper chloride diets whereas no response within copper sulfate diets (P > 0.28).4 Copper source × copper level interaction (P = 0.02). An increasing linear response to increasing copper level (P = 0.02) within

tribasic copper chloride diets whereas a decreasing linear response within copper sulfate diets (P = 0.05).5 Tendency of linear response to copper level (P = 0.12).6 Because 1 piglet was sacrificed at birth for the 3rd and 4th litters, adjustment of excluding the sacrificed piglets was applied

on live born litter size and litter weight that was used in the calculations of survival rate, litter gain, and piglet gain.

24

Table 3. Effects of dietary copper source and level on apparent total tract digestibility (%) during lactation1

Copper source: Tribasic copper chloride Copper sulfate


Item Diet No.: 1 2 3 4 5 6 Source LevelNo. of observations 6 12 10 9 7 6

Dry matter2 87.17 88.15 87.51 86.41 86.79 87.65 0.36 0.03 0.09Nitrogen3 87.90 89.25 88.16 87.22 87.36 87.96 0.54 0.04 0.38Gross energy4 87.71 88.33 87.82 86.70 87.04 88.16 0.41 0.06 0.19Ether extract 76.61 78.51 73.78 67.23 76.04 79.70 3.57 0.51 0.28

1 Fecal samples were collected from the 2nd and 3rd parity sows during d 15 to 17 of lactation.2 Linear response to copper level (P = 0.02).3 Sow parity effect (P < 0.05).4 Tendency of linear response to copper level (P < 0.06).

Table 4. Effects of dietary copper source and level on Htc, Hb, and antioxidant status of lactating sows1, 2


Copper sulfate


Item Diet No.: 1 2 3 4 5 6 Source LevelLactationNo. of observations 8 15 13 13 10 12

Htc, %3 35.58 35.40 34.92 33.29 34.95 34.54 1.04 0.23 0.78Hb, g/dL 11.62 11.92 11.80 11.28 11.53 11.64 0.34 0.30 0.67Total SOD, U/mL3, 4 39.57 43.09 45.77 37.16 39.30 41.74 2.36 0.08 0.09

Cu/Zn SOD, U/mL3, 4 24.90 25.58 30.92 21.29 22.73 26.82 2.05 0.04 0.02Mn SOD, U/mL3, 5 14.67 17.51 14.86 15.87 16.57 14.92 1.14 0.91 0.14

Cp, U/mL5, 6 0.153 0.164 0.130 0.128 0.139 0.117 0.010 0.01 0.02MDA, µM 6.69 6.94 6.54 6.11 7.01 6.12 0.48 0.45 0.32

1 Htc, hematocrit; Hb, hemoglobin; SOD, superoxide dismutase; Cp, ceruloplasmin; MDA, malondialdehyde.2 Blood samples were collected on d 15 of lactation from the 2nd to 4th parity sows.3 Sow parity effect (P < 0.05).4 Linear response to copper level (P < 0.05).5 Quadratic response to copper level (P < 0.05).6 Tendency of linear response to copper level (P < 0.10).

= 0.03) and Zn (P = 0.09) were observed in milk as di-etary Cu level increased. In addition, non-fat solids in milk decreased from sows fed the 20 to 120 mg/kg Cu diets and then increased from the 120 to 220 mg/kg Cu diets (quadratic, P < 0.05).

Tissue Trace Mineral Levels The experimental sows had a greater concentration of Cu (liver and heart) and Zn (liver) when compared to baseline gilts (P < 0.05, Table 6). Moreover, the experi-mental sows tended to have greater concentration of Cu in the kidney than the baseline gilts (P < 0.10). Sows fed the TBCC diets had lower concentrations of Cu (P = 0.04), but greater concentrations of Fe and Mn (P < 0.05) in liver than sows fed the CuSO4 diets. In addition, liver Cu concentration increased with increasing dietary Cu levels (linear and quadratic, P < 0.05). The concentration of Cu in kidney, heart, and ovary was not affected by ei-ther Cu source or level (P > 0.37). Trace mineral concentrations were not affected by sow dietary Cu source or level in various organs of neo-natal piglets (P > 0.20, Table 7). Whereas for weanling

piglets, increasing dietary sow Cu levels resulted in an increase of Cu concentration in liver (linear, P < 0.01, Table 8). However, Zn concentrations in liver tended to decrease as maternal dietary Cu levels increased (linear, P < 0.10).

DiscussionSow and Litter Performance Pigs are Cu tolerant among domestic animal spe-cies (Hill and Spears, 2001) and high concentrations of dietary Cu (100 to 250 mg/kg) are commonly used in nursery and growing-finishing diets as a growth pro-moter. Sows have a much longer lifespan than growing pigs. Thus, one of the concerns of applying pharmaco-logical levels of Cu in sow diets might be the Cu tox-icity due to over-accumulation of Cu in tissues. In the present study, sows fed high Cu diets (120 and 220 mg/kg) for 2.5 yr produced a numerically greater number of litters than sows fed 20 mg/kg Cu (38 and 37 vs. 32 litters; Table 1). This demonstrates that feeding high Cu to sows long-term may not induce any negative effect

25

on fertility of sows. It is in agreement with the results of Cromwell et al. (1993), who reported that there was no difference in the number of litters produced by sows fed diets containing 0 or 250 mg/kg of supplemental Cu (above the basal NRC level in the diets) as CuSO4 for 6 parities. In the present study, increasing dietary Cu levels did not affect litter size of total born, live born, and weaning piglets (P > 0.66; Table 2). Cromwell et al. (1993) previ-ously reported a significant improvement of litter size of total born for sows fed 250 mg/kg supplemented diets compared to sows fed diets without Cu supplementa-tion (P < 0.10). The increasing dietary Cu levels linearly increased piglet weight of total born (P = 0.12) and live born (P = 0.06) piglets; meanwhile, maternal liver Cu concentrations also exhibited a linear increase with in-

creasing dietary Cu levels (P < 0.0001; Table 6). These results might indicate an association between fetal de-velopment and maternal Cu status. This speculation is in accordance with a human study that demonstrated Cu concentrations in the placenta from 20 to 40 yr old mothers at 37 to 41 wk of gestation were positively correlated with neonate weight (Özdemir et al., 2009). Significant interactions between dietary Cu source and level were observed on adjusted piglet weight at wean-ing (P = 0.06) and adjusted piglet weight gain (P = 0.02), which showed increasing linear response within TBCC diets with no response or decreasing linear response within CuSO4 diets, to the increasing dietary Cu lev-els. Tribasic copper chloride has been demonstrated to be a less prooxidative form of Cu than CuSO4 in the intestinal lumen of animals and feed. Studies showed

Table 5. Effects of dietary copper source and level on nutrient concentrations in milk (as-is basis)1


Copper sulfate



Fat, %2 5.72 5.78 6.19 4.98 5.06 6.03 0.38 0.10 0.07Protein, % 4.60 4.62 4.75 4.44 4.19 4.57 0.12 0.02 0.11Lactose, %2 5.95 5.95 5.93 6.16 6.00 5.93 0.06 0.10 0.10Gross energy, Mcal/kg3 1.03 1.03 1.08 0.96 0.94 1.05 0.04 0.07 0.06Total solids, %3 17.26 17.31 17.93 16.50 16.22 17.57 0.45 0.06 0.05Non-fat solids, %4 10.97 10.90 11.08 10.95 10.61 10.90 0.12 0.11 0.13Minerals, µg/mL5

Copper2 1.12 1.23 1.34 1.01 1.12 1.36 0.05 0.13 < 0.0001Iron 2.05 1.66 1.51 1.38 1.63 1.48 0.23 0.22 0.62Zinc2 4.41 3.83 3.74 4.20 3.33 3.49 0.39 0.32 0.17

1 Milk samples were collected from the 3rd and 4th parity sows.2 Linear response to copper level (P < 0.05).3 Tendency of linear response to copper level (P < 0.10).4 Quadratic response to copper level (P < 0.05).5 Manganese was below the detectable level with atomic absorption spectrophotometer.

Table 6. Effects of dietary copper source and level (mg/kg) on tissue trace mineral concentration (DM basis, mg/kg) of sows1, 2

Copper source: Tribasic copper chloride Copper sulfateBaseline

gilts SEMP valuesCopper level, mg/kg: 20 120 220 20 120 220

Item Diet No.: 1 2 3 4 5 6 Source LevelNo. of observations 3 6 4 4 4 4 6

LiverCopper3, 4, 5 123.9 232.5 923.0 298.0 459.8 1372.5 72.1 129.8 0.04 < 0.0001Iron 1363.9 1416.6 979.6 934.0 993.9 852.1 1156.1 159.5 0.04 0.41Manganese3 8.29 8.07 7.03 6.82 6.84 6.73 6.12 0.58 0.04 0.14Zinc3 260.1 234.1 226.4 255.0 236.3 282.9 165.0 27.1 0.46 0.58

CopperKidney6 46.62 38.89 40.73 39.51 36.91 52.71 23.14 11.36 0.97 0.98Heart3 11.51 11.47 11.91 11.58 12.10 14.05 9.82 1.06 0.37 0.43Ovary 9.88 7.62 10.15 7.69 7.27 8.56 5.34 2.18 0.47 0.99

1 A single degree of freedom contrast was conducted to compare experimental sows vs. baseline gilts.2 Tissue samples were collected from sows that completed the 3rd and 4th parity.3 Difference for the contrast of experimental sows vs. baseline gilts (P < 0.05).4 Linear response to copper level (P < 0.0001).5 Quadratic response to copper level (P = 0.04).6 Tendency of difference between experimental sows vs. baseline gilts (P < 0.10).

26

that high dietary levels of CuSO4 may result in elevated MDA concentration in duodenum of nursery pigs and decreased liver vitamin E level in broilers as compared to the same level of Cu in the form of TBCC (Fry et al., 2012; Huang et al., 2015; Luo et al., 2005). In addition, the prooxidant activity exerted by CuSO4 had greater detrimental effects on dietary vitamin activity during storage when compared to TBCC (Lu et al., 2010; Miles et al., 1998).

Apparent Total Tract Digestibility Lactating sows fed TBCC diets had greater ATTD of DM, nitrogen, and GE than those fed CuSO4 diets. It might suggest that TBCC fed sows absorbed more nutrients and consequently provide more nutrients to their progenies through milk. This is consistent with the results of the present study that piglets from TBCC fed sows had greater weaning weight and weight gain dur-ing lactation than those from CuSO4 fed sows.

Antioxidant Status The activity of total and Cu/Zn SOD in sow serum increased linearly with increasing dietary Cu levels re-gardless of Cu sources (P < 0.05). Copper is a critical constituent of Cu/Zn SOD; it is reversibly oxidized and reduced by successive encounters with to yield O2 and H2O2 during SOD catalysis (Tainer et al., 1983). It has been reported that 50 and 250 mg/kg of supplemental Cu as CuSO4 increased serum and erythrocyte SOD activity of growing pigs, respectively, when compared with pigs fed diets without supplemental Cu (Feng et al., 2007; Gonzales-Eguia et al., 2009). Moreover, it also has been reported that greater dietary Cu may lead to elevated SOD activity in other species including beef cattle (40 vs. 0 mg/kg supplemental Cu) and broiler chickens (50 vs. 0 mg/kg supplemental Cu) (Song et al., 2011; Correa et al., 2014).

Table 7. Effects of dietary copper source and level on tissue trace mineral concentration (DM basis, mg/kg) of pig-lets at birth1


Copper sulfate



LiverCopper 156.8 174.4 182.2 154.2 147.4 188.7 20.5 0.65 0.29Iron 496.3 793.7 698.7 675.7 571.4 778.1 144.7 0.92 0.58Manganese 5.62 5.75 6.05 5.14 6.12 6.07 0.43 0.93 0.27Zinc 357.4 412.7 431.8 406.8 390.4 409.9 61.9 0.97 0.82

CopperKidney2 11.85 10.69 11.30 12.36 10.15 13.57 1.18 0.45 0.20Heart 10.17 9.92 9.93 10.04 9.96 10.14 0.19 0.80 0.71

1 Tissue samples were collected from piglets in the 3rd and 4th parity litters.2 Tendency of quadratic response to copper level (P = 0.08).

Table 8. Effects of dietary copper source and level on tissue trace mineral concentration (DM basis, mg/kg) of piglets at weaning1


Copper sulfate



LiverCopper2 358.1 399.3 537.1 379.7 444.9 541.6 27.0 0.29 < 0.0001Iron 152.3 343.3 224.1 299.0 207.6 417.5 80.4 0.32 0.46Manganese 9.69 9.18 10.41 10.55 9.45 9.83 0.67 0.74 0.41Zinc3 391.8 280.3 322.3 384.4 356.3 292.1 42.7 0.72 0.15

CopperKidney 29.86 22.06 29.00 32.47 28.45 32.77 4.57 0.27 0.38Heart4 8.68 9.06 9.04 8.78 9.17 8.98 0.22 0.79 0.25

1 Tissue samples were collected from piglets in the 3rd and 4th parity litters.2 Linear response to copper level (P < 0.0001).3 Tendency of linear response to copper level (P = 0.06).4 Sow parity effects (P < 0.05).

27

Milk Composition In the current study, the improved milk composition of sows fed TBCC diets compared to those fed CuSO4 diets may explain the greater adjusted piglet weight at weaning, as well as the greater adjusted litter and piglet weight gain of litters from TBCC fed sows. In addition, the greater nutrient concentrations in milk might also indicate an improved mammary gland development. Mammary gland development occurs throughout many stages of growth and reproduction in swine, and various hormones are involved in the control of mammary de-velopment (Farmer and Hurley, 2015). Growth hormone (GH), which is secreted by the pituitary gland, regulates the development of mammary gland mainly through in-sulin-like growth factor-1 (IGF-1) (Ruan and Kleinberg, 1999; Gallego et al., 2001). It has been demonstrated by many studies that high dietary Cu levels increased ex-pression of GH and growth hormone-releasing hormone mRNA, as well as serum IGF-1 levels in growing pigs (Wang et al., 2016; Yang et al., 2011; Zhou et al., 1994). Therefore, it is speculated that the greater nutrient levels in milk produced by TBCC fed sows may be associated with the Cu-induced changes of hormone secretion.

Tissue Trace Mineral Levels Trace mineral concentrations of neonatal piglets did not differ across treatments. This indicates that transfer of trace minerals from the maternal tissues to fetuses was not affected by dietary Cu source and level. How-ever, liver Cu concentrations of weanling piglets linearly increased as sow dietary Cu level increased (P < 0.05). This is associated with Cu concentrations in milk, which exhibited a significant linear increase with increasing sow dietary Cu levels. Moreover, since Cu level was concentrated in sow feces (Diet 1 to 6: 296, 964, 1607, 407, 1127, and 1782 mg/kg during gestation; 185, 876, 1559, 252, 961, and 1583 mg/kg during lactation); and the suckling piglets were able to access sow feces during lactation, the greater liver Cu accumulation of weanling piglets might also be partially attributed to coprophagy.

Conclusions In the current study, sows fed with TBCC diets had improved nutrient digestibility, lower Cu but greater Fe and Mn concentrations in liver, higher nutrient levels in milk, greater serum antioxidant enzyme activity during lactation, and greater litter performance, when com-pared to sows fed with CuSO4 diets. Additionally, in-creasing dietary Cu levels linearly increased digestibility

of DM and GE during lactation, the antioxidant enzyme activity of sows, nutrient levels in milk, total born and live born piglet weight, and liver Cu concentrations in sows and weanling piglets. Results of the present study indicate that TBCC might be a superior Cu source com-pared to CuSO4 for sows; the greater dietary Cu levels is beneficial to improve piglet birth weight, serum SOD activity, and Cu concentration in tissue of sows and pig-lets without showing any apparent negative effects on sows and progenies.

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Paiva, S. da Luz e Silva, and A. S. Netto. 2014. Effects of supplementation with two sources and two levels of copper on meat lipid oxidation, meat colour and superoxide dismutase and glutathione peroxidase enzyme activities in Nellore beef cattle. Br. J. Nutr. 112: 1266-1273. doi:10.1017/s0007114514002025

Cromwell, G. L. 2001. Antimicrobial and promicrobial agents. In: A. J. Lewis and L. L. Southern, editors, Swine Nutrition. 2nd ed. CRC Press, New York, NY. p. 398.

Cromwell, G. L., H. J. Monegue, and T. S. Stahly. 1993. Long-term effects of feeding a high copper diet to sows during gestation and lactation. J. Anim. Sci. 71: 2996-3002. doi:10.2527/1993.71112996x

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Feng, J., W. Q. Ma, Z. L. Gu, Y. Z. Wang, and J. X. Liu. 2007. Effects of dietary copper (II) sulfate and copper pro-teinate on performance and blood indexes of copper status in growing pigs. Biol. Trace Elem. Res. 120: 171-178. doi:10.1007/s12011-007-8001-y

Fry, R. S., M. S. Ashwell, K. E. Lloyd, A. T. O’Nan, W. L. Flowers, K. R. Stewart, and J. W. Spears. 2012. Amount and source of dietary copper affects small intestine morphology, duodenal lipid peroxidation, hepatic oxi-dative stress, and mRNA expression of hepatic copper regulatory proteins in weanling pigs. J. Anim. Sci. 90: 3112-3119. doi:10.2527/jas2011-4403

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29

Development and Nutritional Value of Advanced Soybean Products Used in Diets

for Young PigsLaia Blavi and Hans H. Stein

Department of Animal Sciences University of Illinois, Urbana 61801


SummarySoybean meal is an excellent source of protein for pigs because of its excellent amino acid profile and its high amino acid digestibility. However, the presence of anti-nutritional factors such as trypsin inhibitors, lectins, oligosaccharides, and antigenic proteins reduces the inclusion level in weanling diets. The oligosaccharides can be reduced or eliminated by processing the soybean meal via aque-ous ethanol extraction, fermentation, or enzymatic treatment. Reducing the concentration of oligo-saccharides increases the concentration of other nutrients such as crude protein. Soy protein concen-trate and soy protein isolate have low levels of stachyose and raffinose and antigenic proteins, and greater digestibility of amino acids and energy than soybean meal. Enzyme-treated soybean meal also has low concentration of oligosaccharides, and fermented soybean meal contains no stachyose or raffinose. However, fermented soybean meal sometimes has a reduced digestibility of lysine be-cause of Mailard reactions caused by excess heating during drying after fermentation, but this is usually not observed in enzyme-treated soybean meal where the amino acid digestibility is similar to soybean meal.

Introduction Soybeans is one of the most important crops in the U.S. and the co-product soybean meal (SBM) is the pri-mary source of protein in swine diets because of its fa-vorable concentration and balance of digestible amino acids (AA). Domestic livestock and poultry consumed 28 million metric tons of SBM in 2010, using nearly 80% of all the soybean meal processed in the U.S. However, raw soybeans contain anti-nutritional factors: trypsin inhibitors, lectins, phytic acid, oligosaccharides (raffi-nose, stachyose, and verbascose), and antigenic protein (glycinin and β-conglycinin; Baker, 2000; NRC, 2012; He et al., 2015). To reduce the concentration of some of the anti-nutritional factors such as trypsin inhibitors, soybean products need to be heated before being fed to swine because trypsin inhibitors are heat-labile. Soybeans contain approximately 37% crude protein and 20% fat (Table 1). However, after crushing, most of the fat is removed via solvent extraction, and the result-ing SBM contains less than 2% fat. The concentration of total carbohydrates in intact soybeans is 35 to 40% with approximately 15% being non-structural carbohydrates, such as sucrose, uronic acid, oligosaccharides, and free

sugars. The concentration of oligosaccharides (raffinose, stachyose, and verbascose) in soybeans is between 4 and 7% (NRC, 2012). However, in SBM the concentration of oligosaccharides may be between 7 and 11%, where the concentration of stachyose is between 5.1 and 7.3% and the concentration of raffinose is between 1.1 and 3.8% (Cervantes-Pahm and Stein, 2010; NRC, 2012). The main oligosaccharides in soybeans are raffinose, stachyose, and verbascose, and they are commonly called α-galactosides because of the presence of galac-

Table 1. Nutritional composition of U.S. soybeans and U.S. soybean meal (NRC, 2012).

Full-fat soybeans

48% U.S. SBM, dehulled

Dry matter, % 92.36 89.98Crude protein, % 37.56 47.73Ash, % 4.89 6.27Ether extract, % 20.18 1.52Carbohydrates, % Sucrose 6.42 4.30 Raffinose 0.77 3.78 Stachyose 3.89 7.33 Verbascose 0.03 0.00 Starch 1.89 1.89 NDF 10.00 8.21

30

tose in the structure of these three oligosaccharides. Raffinose consists of one unit of glucose, one unit of fructose, and one unit of galactose. Stachyose and ver-bascose have a structure that is similar to raffinose with the exception that they contain two or three units of ga-lactose, respectively. The glucose and fructose units in the oligosaccharides are connected by an α-(1-2) glyco-sidic linkage, whereas an α-(1-6) linkage connects glu-cose to galactose and also connects the galactose units in stachyose and verbascose. Therefore, the glycosidic linkages in the α-galactosides may be hydrolyzed by the enzyme α-galactosidase. However, pigs do not secrete the digestive enzymes necessary to cleave α-1,6 linkages and raffinose, stachyose, and verbascose are, therefore, considered indigestible by pigs, and are fermented in the digestive tract (Canibe and Bach Knudsen, 1997). How-ever, it has been demonstrated that the ileal digestibility of α-galactosides is between 50 and 80% (Bengala-Freire et al., 1991; Canibe and Bach Knudsen, 1997; Smiricky et al., 2002), which indicates that there is considerable fermentation taking place in the small intestine of pigs. This fermentation allows pigs above approximately 20 kg to utilize the energy from the oligosaccharides in the form of short chain fatty acids. However, younger pigs fed large quantities of SBM do not handle the fer-mentation of these oligosaccharides very efficiently, and negative side effects such as diarrhea, gastrointestinal discomfort, and a reduction of weight gain are observed (Liying et al., 2003). Inclusion of SBM in diets fed to weanling pigs is, therefore, usually restricted to less than 20%. The total tract digestibility is considered to be 100% because any α-galactosides that are not fer-mented in the small intestine are rapidly fermented in the large in-testine.

Removal of the Oligosaccharides Removing the oligosaccharides from conventional SBM may be achieved by removing the non-protein constituents from dehulled and deffated soybeans, by fermen-tation or by enzyme treatment. Therefore, different products can be obtained: soy protein concen-trate (SPC), soy protein isolate (SPI), fermented soybean meal (FSBM), or enzyme-treated soy-bean meal (ESBM).

Soy Protein Concentrate Soy protein concentrate by definition contains a minimum of 65% crude protein (CP) on a dry matter (DM) basis (Endres, 2001), and it is produced by acid leaching, extraction with aqueous alcohol, or by dena-turing the protein with moist heat before extraction with water (Endres, 2001). Thus, SPC contains fewer trypsin inhibitors, sucrose, raffinose, and stachyose than SBM (Oliveira and Stein, 2016), and the concentration of CP and AA are greater than in SBM (Table 2). The AA digestibility in SPC is similar to SBM, except for some AA where the standardized ileal digestibility (SID) is greater in SPC than in SBM (Oliveira and Stein, 2016; Pedersen et al., 2016). However, SPC contains more DE and ME than SBM (Oliveira and Stein, 2016). Reduction in particle size of SBM improves the di-gestibility of most indispensable AA (Fastinger and Ma-han, 2003) and the values of DE and ME (Rojas and Stein, 2015). Therefore, when SPC is ground to 70 or 180 µm the SID of arginine, isoleucine, phenylalanine, and tryp-tophan is greater than in SBM. However, there are no dif-ferences among conventional SBM and SPC ground to 70, 180, or 700 µm in DE and ME (Casas et al., 2017). Addition of SPC to weanling pig diets at the ex-pense of animal proteins does not affect the growth performance during the initial 4 weeks post-weaning (Guzmán et al., 2016; Casas and Stein, 2017), but SPC re-duces the incidence of post-weaning diarrhea (Guzmán et al., 2016). Therefore, SPC may be used in diets fed to weanling pigs as a replacement for animal proteins.

Table 2. Nutrient composition of soybean meal, soy protein concentrate, and soy protein isolate (NRC, 2012).

Soybean Meal

Soy Protein Concentrate

Soy Protein Isolate

Fermented SBM

Enzyme-treated

SBMDry matter, % 89.9 92.6 93.7 92.7 92.9Crude protein, % 47.7 65.2 84.8 55.6 54.1Ether extract, % 1.5 1.1 2.8 1.8 2.3Ash, % 6.3 6.11 4.2 7.1 7.0Carbohydrates, %

Sucrose 4.30 0.67 0.13 - -Raffinose 3.78 0.46 - - -Stachyose 7.33 0.91 - - -Verbascose 0.00 - - - -

Amino Acids, %Arginine 3.45 4.75 6.14 3.95 3.70Histidine 1.28 1.70 2.19 1.41 1.37Isoleucine 2.14 2.99 3.83 2.48 2.55Leucine 3.62 5.16 6.76 4.09 4.25Lysine 2.96 4.09 5.19 3.20 3.14Methionine 0.66 0.87 1.11 0.71 0.75Phenylalanine 2.40 3.38 4.40 2.78 2.87Threonine 1.86 2.52 3.09 2.13 2.09Tryptophan 0.66 0.81 1.13 0.72 0.69Valine 2.23 3.14 4.02 2.57 2.67

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Soy Protein Isolate Soy protein isolate contains at least 80% CP (Mid-delbos and Fahey, 2008). It is produced by solubilizing the protein at neutral and slightly alkaline pH, and the extract is then precipitated by acidification to obtain the protein isolate (Berk, 1992). Therefore, most of the non-protein constituents from soybeans are removed, and SPC therefore contains very few trypsin inhibi-tors, limited fiber, and practically none of the oligosac-charides (Table 2). The allergenic proteins glycinin and β-conglycinin are deactivated in soy protein isolate, and also in SPC because they are produced by extraction at temperatures greater than 50°C (Sissons et al., 1982). The AA digestibility of SPI is similar to that in casein (Cervantes-Pahm and Stein, 2010) and similar to SBM, but for some AA, SPI has greater SID values than SBM (Pedersen et al., 2016). Soy protein isolate is well toler-ated by weanling pigs (Li et al., 1991) but its high cost of production makes it uncommon in commercial pig feed production.

Fermented Soybean Meal Fermented soybean meal is produced by inoculat-ing conventional soybean meal with the bacterium As-pergillus oryzae or other microbes (Hong et al., 2004). Raw soybeans are soaked in distilled water for 3 hours and placed in an autoclave at 100–120°C for 20 min-utes. After that, autoclaved soybeans are cooled to room temperature for 3 hours. The soybeans are then inocu-lated with A. oryzae and placed in an incubator for 48 hours at 30°C with 90% moisture. After fermentation, soybeans are dried at 50–60°C and ground in a hammer mill (Hong et al., 2004). Fermented soybean meal contains more DM, CP, and ash than conventional SBM (Table 2). The absence of sucrose, stachyose, and raffinose in FSBM is attrib-uted to the production of α-galactosidase by Aspergillus oryzae during the fermentation process (Cervantes-Pahm and Stein, 2010). The disappearance of these sac-charides is the main reason for the analyzed increase in the concentration of other nutrients in FSBM as com-pared with SBM. Yoon (2012) analyzed 4 different samples of FBSM and observed that the concentration of CP is between 53 and 58%, which is greater than in conventional SBM. However, the SID of lysine is lower in FSBM than in SBM (Cervantes-Pahm and Stein, 2010) and lower lysine-to-CP ratio compared with SBM, SPC, SPI, and ESBM (Pedersen et al., 2016). This is likely a result of the heat that is used during drying of FSBM, which may

result in heat damage. Heat damage may result in Mail-lard reactions that can destroy some of the lysine in the FSBM (Stein et al., 2009), and Maillard reactions can also result in reduced SID of lysine (Stein et al., 2009). Peptide size distribution does not differ between SBM and FSBM and there is no evidence for hydrolysis of the peptides in FSBM (Cervantes-Pahm and Stein, 2010). The concentration of glycinin and β-conglycinin in FSBM is similar to SBM, which is in contrast with SPI that has low concentration of the allergenic proteins (Cervantes-Pahm and Stein, 2010). Antigenic proteins glycin and β-conglycinin reduce ADG and G:F in young pigs (Zhao et al., 1998) and may reduce villus height in the small intestine and decrease nitrogen digestibility in pigs (Li et al., 1991).

Enzyme-treated Soybean Meal Enzyme-treated SBM is produced by treating de-hulled solvent-extracted SBM for several hours with a proprietary blend of enzymes (Goebel and Stein, 2011). Enzyme treatment removes sucrose and reduces the concentrations of oligosaccharides and allergenic pro-teins (Cervantes-Pahm and Stein, 2010). Therefore, the concentration of CP and other nutrients is greater in ESBM than in SBM (Table 2). In addition, there are no differences in SID of AA between SBM and ESBM (Cervantes-Pahm and Stein, 2010; Pedersen et al., 2016). Several studies have demonstrated that ESBM is well accepted by young pigs, and ESBM may, therefore, replace animal proteins in starter diets for pigs.

Conclusion The reduction of oligosaccharides and antigenic proteins in SPC, SPI, FSBM and ESBM increases the nutritional value to young pigs and the AA digestibility is usually similar to that in SBM or greater. As a result, these special soybean products may be used in diets fed to weanling pigs as a replacement for fishmeal or others animal proteins.

ReferencesBaker, D. H. 2000. Nutritional constraints to the use of

soy products by animals. Pages 1-12 in Soy in Ani-mal Nutrition. J. K. Drackley, ed. Fed. Anim. Sci. Soc., Champaign, IL.

Bengala-Freire, J., A. Aumaitre, and J. Peiniau. 1991. Ef-fects of feeding raw and extruded peas on ileal diget-ibility, pancreatic enzymes and plasma glucose and insulin in early weaned pigs. J. Anim. Phys. Anim. Nutr. 65:154–164. doi:10.1111/j.1439-0396.1991.tb00253.x

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Berk, Z. 1992. Technology of production of edible flours and protein products from soybeans. Rome: Food and Agriculture Organization of the United Na-tions. p. 133.

Canibe, N., and K. E. Bach Knudsen. 1997. Digestibil-ity of dried and toasted peas in pigs. 1. Ileal and total tract digestibilities of carbohydrates. Anim. Feed Sci. Technol. 64:293–310. doi:10.1016/S0377-8401(96)01032-2

Casas, G. A., C. Huang, and H. H. Stein. 2017. Nutri-tional value of soy protein concentrate ground to different particle sizes and fed to pigs. J. Anim. Sci. 95:827-836. doi:10.2527/jas2016.1083

Cervantes-Pahm, S. K., and H. H. Stein. 2010. Ileal di-gestibility of aminoacids in conventional, ferment-ed, and enzyme-treated soybean meal and in soy protein isolate, fish meal, and casein fed to wean-ling pigs. J. Anim. Sci. 88:2674-2683. doi:10.2527/jas.2009-2677.

Endres, J. G. 2001. Soy protein products: characteris-tics, nutritional aspects, and utilization. Urbana, IL: American Oil Chemists’ Society.

Fastinger, N. D., and D. C. Mahan. 2003. Effect of soy-bean meal particle size on amino acid and energy di-gestibility in growing pigs. J. Anim. Sci. 81:697-704. doi:10.2527/2003.813697x

Goebel, K. P., and H. H. Stein. 2011. Phosphorus digest-ibility and energy concentration of enzyme treated and conventional soybean meal fed to weanling pigs. J. Anim. Sci. 89:764-772. doi:10.2527/jas.2010-3253

Guzmán, P., B. Saldaña, L. Cámara, and G. G. Mateos. 2016. Influence of soybean protein source on growth performance and nutrient digestibility of piglets from 21 to 57 days of age. Anim. Feed Sci. Technol. 222:75-86. doi:10.1016/j.anifeedsci.2016.10.004

He, L., M. Han, S. Qiao, P. He, D. Li, and X. Ma. 2015. Soybean antigen proteins and their intestinal sensi-tization activities. Curr. Protein Pept. Sci. 16:613-21. doi:10.2174/1389203716666150630134602

Hong, K.J., C. H. Lee, and S. W. Kim. 2004. Aspergillus oryzae GB-107 fermentation improves nutritional quality of food soybeans and feed soybean meals. J. Med. Food. 7:430-435. doi:10.1089/jmf.2004.7.430

Li, D. F., J. L. Nelssen, P. G. Reddy, F. Blecha, R. D. Kl-emm, D. W. Giesting, J. D. Hancock, G. L. Allee, and R. D. Goodband. 1991. Measuring suitability of soybean products for early-weaned pigs with im-munological criteria. J. Anim. Sci. 69:3299–3307. doi:10.2527/1991.6983299x

Liying, Z., D. Li, S. Qiao, E. Johnson, B. Li, P. Thacker, and I. K. Han. 2003. Effects of stachyose on per-formance incidence and intestinal bacteria in weanling pigs. Arch. Anim. Nutr. 57:1-10. doi: 10.1080/0003942031000086662

NRC. 2012. Nutrient requirements of swine. 11th ed. National Academy Press, Washington DC.

Middelbos, I. S., and G. C. Fahey. 2008. Soybean carbo-hydrates. In: L. A. Johnson, P. J. White, and R. Gallo-way, editors, Soybeans: chemistry, production, pro-cessing, and utilization. Academic Press and AOCS Press. p. 269-296. doi:10.1016/B978-1-893997-64-6.50012-3

Oliveira, M. S., and H. H. Stein. 2016. Digestibility of energy, amino acids, and phosphorus in a novel source of soy protein concentrate and in soybean meal fed to growing pigs. J. Anim. Sci. 94:3343-3352. doi:10.2527/jas2016-0505.

Pedersen, C., J. S. Almeida, and H. H. Stein. 2016. Analy-sis of published data for standardized ileal digestibil-ity of protein and amino acids in soy protein fed to pigs. J. Anim. Sci. 94:340-343. doi:10.2527/jas2015-9864

Rojas, O. J., and H. H. Stein. 2015. Effects of reducing the particle size of corn grain on the concentration of di-gestible and metabolizable energy and on the digest-ibility of energy and nutrients in corn grain fed to growing pigs. Livest. Sci. 181:187-193. doi:10.1016/j.livsci.2015.09.013

Sissons, J. W., A. Nyrup, P. J. Kilshaw, and R. H. Smith. 1982. Ethanol denaturation of soya bean protein an-tigens. J. Sci. Food Agric. 33:706-710. doi:10.1002/jsfa.2740330804

Smiricky, M. R., C. M. Grieshop, D. M. Albin, J. E. Wub-ben, V. M. Gabert, and G. C. Fahey, Jr. 2002. The influence of soy oligosaccharides on apparent and true ileal amino acid digestibilities and fecal consis-tency in growing pigs. J. Anim. Sci. 80:2433–2441. doi:10.2527/2002.8092433x

Stein, H. H., S. P. Connot, and C. Pedersen. 2009. Energy and nutrient digestibility in four sources of distill-ers dried grains with solubles produced from corn grown within a narrow gragraphical area and fed to growing pigs. Asian-asutralas. J. Anim. Sci. 22:1016-1025. doi:10.5713/ajas.2009.80484

Yoon, J. 2012.Effects of genetic selection and processing of soybeans and soybean meal on nutritional quality. Masters Thesis. Univ. Illinois, Urbana-Champaign.

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Nutritional Regulation of Piglet Gut HealthSheila K. Jacobi, Emily E. Chucta, and Ariel Taylor

Department of Animal Sciences Ohio State University, Columbus, OH 43210


Jack Odle Department of Animal Sciences

North Carolina State University, Raleigh, NC 27695

SummaryGastrointestinal health issues rank among the highest causes of neonatal morbidity and mortality across most mammalian species, including pigs (USDA, 2012). Gut health is an area of research with significant impact to swine nutrition because without a healthy intestinal tract there are se-vere impacts from poor nutrient digestion and absorption and slow growth to increased intestinal pathogens which can result in death. However, the definition of gut health is vague without a precise etiology. Nevertheless, it is known that nutrition has a great impact on gut development. The stresses of production, growth, and changing nutrition over the growth period often present a challenge to optimum intestinal health, and therefore, efficient growth for optimal least cost swine production. For years, animal livestock has utilized antimicrobial growth promotants to mitigate some of the challenges of gut health and to optimize efficient growth. However, with pressure to reduce the feed-ing of antimicrobial growth promotants in livestock production, it has forced the industry to look at alternative ways to optimize growth performance while minimizing use of antimicrobials. The development of new/different management and feeding strategies to stimulate gut development and a healthy microbiome in the neonate is necessary, and understanding how nutritional bioactive compounds promote gut and microbiome development in early life is necessary for optimizing gut health and immune function throughout production.

Introduction The definition of gut health is more than a homeo-static gastrointestinal tract. Bischoff (2011) defined gut health by five major criteria: 1. the effective digestion and absorption of food, 2. absence of GI illness, 3. a nor-mal and stable microbiome, 4. effective immune status, and 5. status of well-being. This definition of gut health does not only associate health with pathogenic disease disruption of intestinal health, but allows for inclusion of the concepts of how prevention and avoidance of gastrointestinal disease are part of the gut health equa-tion. For the swine industry we know many stressors in-volved in production can disturb pig performance, and it is important to understand how developing nutrition-al and management programs may enhance gut health through prevention and/or treatment to optimize pig performance. One of the five criteria that Bischoff (2011) defines as a part of gut health is a normal and stable microbi-ome. In recent years, the focus on how microbiome affects the overall health of humans and animals has

become a focus of the nutritional community because diet significantly impacts the gut microbiome (Isaacson and Kim, 2012). Intestinal microbiota are acquired early in life and play many roles in host’s health. They affect maturation of the gut, program/mature the developing immune system, competitively exclude pathogenic or-ganisms that cause disease, and the microbes are criti-cal in nutrient digestion such as fiber and synthesis of vitamins. Additionally, imbalance in a homeostatic microbial population in the gut is known to cause dis-ruption of gut barrier function (Pluske et al., 2018), and gut barrier function is critical in maintenance of the five criteria of gut health. Therefore, optimal nutritional support for the developing intestinal tract, microbiome, and immune system of young pigs is important for ef-ficient growth and health, and likely critical for life-long health of the intestinal track given the impact on muco-sal immune development (Frese et al., 2015). Bioactive components are particularly important in prevention and possibly management of gut health. With new feed supplements being developed to moderate neonatal in-

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testinal issues that improve survivability and improve production performance in swine research on under-standing how nutrients could be used in preventive health are important. Additionally, there are increasing pressures to mitigate intestinal challenges and improve piglet performance without the use of antimicrobial compounds and high mineral feeding that are known to enhance performance, but also have negative environ-mental impacts (Pluske, 2013). Therefore understand-ing how other bioactive compounds may alter microbial populations in the developing piglet may lead to a better understanding of how we use nutrition to optimize gut health and immune status in pigs. Studies of gut microbiota show that early postna-tal environmental exposures play a critical role in de-termining the overall composition of the adult animal and programing of the adult immune system (Kau et al., 2011). Early dietary exposure to mother’s microbiota and nutrients of maternal milk which stimulate micro-bial composition are some of the first exposures of the neonatal GI tract to colonization (Bode, 2012). Of the dietary components known to modulate neonatal gut microbial populations, the oligosaccharide component composition are the most influential. Oligosaccharides, like prebiotics, are undigested by the neonate and have several functions in the development of young piglets

gastrointestinal tract including pathogen binding, im-mune system development, and serving as a growth factor to beneficial bacteria (Tao et al., 2010; Salcedo et al., 2016). In addition to prebiotics developing the mi-crobiome, they also are known to increase short chain fatty acid (SCFA) production by the microbiome. SCFA are produced by gut microbiome members (Nicholson et al., 2012) (e.g., Eubacterium, Roseburia, Faecalibacte-rium, and Coprococcus) directly from sugar to produce a mixture of fatty acids - acetate, propionate, butyrate, isobutyrate, 2-methylpropionate, valerate, isovalerate, and hexanoate (Nicholson et al., 2012). These SCFA’s are associated with decreased colonic pH, inhibition of pathogens; increased water and sodium absorption; and an energy source to colonic epithelial cells. Acetate accumulates in circulation making it a very good can-didate for pre- or probiotic supplementation13. Acetate producing bacteria include Akkermansia muciniphila, Bacteroides, Bifidobacterium, Prevotella, Ruminococcus (Koh et al., 2016). Treatments that increase these organ-isms will result in health benefits for an animal and de-crease pathogens. A second beneficial SCFA is butyrate. Acetate can be converted to butyrate by the microbiome, which is a primary energy source for colonocytes, is anti-inflam-matory, promotes cell function and differentiation (Koh

Figure 1. Principle Coordinate Analysis of Changing Colonic Microbial Taxa in Suckling Piglets. Each plot represents a either trial day 0, 7, 14 or 21. Dietary treatments are PRE = prebiotics; ARA = arachidonic acid; or a combination of PRE+ARA; Supplementa-tion is denoted by + or – in the figure legends on the principle coordinate plots.

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Table 3. Dietary Effects on Fatty Acid Composition (%wt) of Colonic Mucosa

CONT ARA SE P-Value -PRE +PRE SE P-ValueC18:0 21.36 21.32 0.94 0.98 23.65a 19.03b 1.02 0.002C18:1c 23.49 22.54 0.72 0.35 22.37 23.66 0.78 0.21C18:2c n6 14.10a 12.81b 0.43 0.04 11.26b 15.67a 0.46 0.001C18:3 n3 0.40 0.40 0.04 0.99 0.31b 0.48a 0.03 0.001C20:4 n6 3.83a 6.67b 0.33 0.001 3.74b 6.67a 0.37 0.001a,b Means within a row with different superscripts differ (P<0.05).CONT = control, ARA = arachidonic acid, PRE = prebiotics.

Table 1. Dietary Effects on Intestinal pH

CONT PRE ARA PRE + ARA SE P-ValueIleum 7.36 7.29 7.36 7.31 0.05 0.647Cecum 6.22a 5.74b 6.24a 5.84b 0.06 0.001Proximal Colon 6.38a 5.71b 6.34a 5.78b 0.07 0.001Distal Colon 6.50a 6.05b 6.60a 6.15b 0.07 0.001a,b Means within a row with different superscripts differ (P<0.05)Dietary treatments are CONT = control, PRE = prebiotics, ARA = arachidonic acid, or a combination of PRE+ARA.

Table 2. Dietary Effects on VFA Concentrations in Distal Colon

CONT PRE ARA PRE + ARA SE P-ValueAcetate 41.5a 51.4b 44.7a 48.6b 2.2 0.001Propionate 12.3a 18.0b 13.7a 16.2b 1.5 0.001Butyrate 7.2a 8.4b 7.9a 8.4b .3 0.001Total 71.0a 87.4b 76.8a 83.1b 3.3 0.001a,b Means within a row with different superscripts differ (P<0.05).Dietary treatments are CONT = control, PRE = prebiotics, ARA = arachidonic acid, or a combination of PRE+ARA.

et al., 2016) - which also occurs with bacterial assocation, and significantly changes response to inflammation (Kol et al., 2014). Butyrate also regu-lates immune cells responses and tight junctions involved in gut barrier function (Koh et al., 2016; Yan and Ajuwon, 2017). The effects of dietary long-chain polyunsaturated fatty acids (LCP-UFA) on intestinal microbiota have not been as well researched. Neilsen et al. (2007) showed that daily fish oil supplementation for one month in-fluenced the composition of the fecal bacteria in human babies. Similarly, piglets fed formula containing LCP-UFA significantly influenced overall bacterial composition and the size of the Bacteroides community (Anderson et al., 2011). These and others report positive effects of LCPUFA on the numbers of different lactic acid bacteria which have benefit to the developing intestine (Anderson et al., 2011). Therefore, we are interested in how diet not only provides essential nutrients for growth and development, but how indi-vidual nutrients have critical bioactive roles in modu-lating gut physiology. Understanding the interaction of these nutrients are a way for nutritionist to modify dietary programs for optimizing piglet growth and sur-vivability.

Methods Full term crossbred pigs were vaginally delivered and allowed to suckle colostrum for 24 hours and then individually housed and fed milk replacer in an environ-mentally controlled barn. Pigs were allocated, according to body weight and litter origin, to one of four dietary treatments differing in prebiotic and fatty acid compo-sition: 1) formula containing no prebiotic and baseline LCPUFA (CONT), 2) formula enriched with 4g/L GOS + 4g/L PDX (PRE), 3) formula enriched with 2.5% ARA (LCPUFA), or 4) formula enriched with both PRE & LCPUFA (PRE+LCPUFA). They were fed to ~60% ad

libitum with fresh diets offered three times a day for 21 days. Fecal swabs were collected weekly and microbi-ome analysis was performed via 16S rDNA Illumina sequencing. On day 21, pigs were euthanized, intestinal pH and colonic mucosal scrapings and digesta were im-mediately collected and snap frozen for further analysis. Fatty acid composition of colonic mucosa was analyzed via GC-MS and volatile fatty acids composition of the colonic digesta was analayzed via GC-FID. Data were analyzed according to a randomized block design using general linear models procedures of SAS (SAS, Cary, NC). Microbiome were analyzed by using R multivari-ate analysis.

Results Diet did not affect growth (P > 0.1). Microbial taxa significantly changed from d 0 to d 21 in a diet-dependent manner with diets containing prebiotics clustering together by d 7 post-feeding and becoming more closely associated with longer feeding times (P < 0.05). Specifically, an increase in bacterial taxa belong-ing to the phyla Bacteroidetes was observed (P < 0.05). Fatty acid inclusion in the diets significantly changed the fatty acid composition of the colonic mucosa. Co-lonic mucosal arachidonic acid (ARA) percent (ARA)

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(w/w) increased from 3.83 in controls to 6.67 in pigs fed diets enriched in ARA (P < 0.001), with reciprocal re-ductions in linoleate concentration. Dietary prebiotics (PRE) also increased ARA and linoleate concentration but reduced stearate concentration in colonic mucosa (P < 0.002). Additionally, dietary PRE reduced colonic pH associated with increased concentration of VFAs (P < 0.05). In conclusion, addition of PRE to milk replacer progressively alters microbial taxa over the first three weeks of life and increases colonic fermentation. Di-etary ARA has minimal impact on microbial taxa but enriches colonic mucosa.

Conclusions The aim of the study was to understand the impact of dietary fatty acids and prebiotics on changes in in-testinal microbial populations that may have beneficial outcomes on piglet gut health. To investigate this goal we fed one-hundred and two crossbred day old piglet on milk replace supplemented with prebiotics or LCP-UFA for 21 days. Additionally, the interaction of bioac-tive nutrients was also investigated by including a treat-ment with a combination of prebiotic plus LCPUFA. Diets containing dietary prebiotics had the most signifi-cant effects on gut microbial population of the devel-oping neonate. Microbial populations of piglets being fed prebiotics had a progressive shift in microbial taxa over the first three weeks of life. The early instability of the microbial population in the gut lends itself to a time when diet could have significant impact on microbial communities and could impact long-term intestinal mi-crobe populations and modify intestinal health. Piglets fed prebiotics also had increased total volatile fatty acids in the colonic digesta due to an increase in the individ-ual VFAs, actetate, propionate, and butyrate at the end of the study. The decrease in colonic digesta pH caused by changes in microbial populations, and the produc-tion of VFA are associated with competitive exclusion of pathogenic microorganism (Nicholson et al., 2012; Koh et al., 2016). Arachidonic acid did not significantly change micro-bial composition of the piglet intestine, but did modify the fatty acid composition of cell membranes. Further analysis will need to be completed to understand what impact this could have on gut health. However, previ-ously we have shown metabolites of long-chain poly-unsaturated fatty acids have significant impact on gut barrier function following an intestinal insult (Jacobi et al., 2011). Modifying the gut environment during development using nutritional interventions may have significant impact on gut health during challenges later

in swine production. Although long term studies would need to be completed in swine to truly understand the impact of early neonatal nutrition in programming gut microbial population that affect overall intestinal health in later production stages, it seems reasonable there could be a significant impact to improve swine gut health.

ReferencesAndersen, A. D., L. Molbak, T. Thymann, K. F. Mi-

chaelsen, and L. Lauritzen. 2011. Dietary long-chain n-3 PUFA, gut microbiota and fat mass in early post-natal piglet development--exploring a potential in-terplay. Prostaglandins Leukot. Essent. Fatty Acids. 85:345-351.

Bischoff, S.C. 2011. ‘Gut health’: a new objective in med-icine? BMC Med. 14;9:24. doi: 10.1186/1741-7015-9-24.

Bode, L. 2012. Human milk oligosaccharides: every baby needs a sugar mama. Glycobiology. 22:1147-1162.

Frese, S. A., K. Parker, C. C. Calvert, and D. A. Mills. 2015. Diet shapes the gut microbiome of pigs during nursing and weaning. Microbiome. 3:28-38.

Isaacson, R., and H. B. Kim. 2012. The intestinal micro-biome of the pig. Animal Health Research Reviews 13(1); 100–109.

Jacobi, S.K., and J. Odle. 2012. Nutritional factors influ-encing intestinal health of the neonate. Adv. Nutr. 1;3(5):687-96. doi: 10.3945/an.112.002683.

Jacobi, S. K., A. J. Moeser, B. A. Corl, R. J. Harrell, A. T. Blikslager, and J. Odle. 2012. Dietary long-chain PUFA enhance acute repair of ischemia-injured in-testine of suckling pigs. J. Nutr. 142(7):1266-1271.

Kau, A. L., P. P. Ahern, N. W. Griffin, A. L. Goodman, and J. L. Gordon. 2011. Human nutrition, the gut microbiome and the immune system. Nature. 474:327-336.

Koh, A.; F. De Vadder, P. Kovatcheva-Datchary, and F. Backhed. 2016. From dietary fiber to host Physiol-ogy: short-chain fatty acids as key bacterial metabo-lites. Cell. 165 (6), 1332-1345.

Kol, A., S. Foutouhi, N. J. Walker, N. T. Kong, B. C. Weimer, and D. L. Borjesson. 2014. Gastrointesti-nal microbes interact with canine adipose-derived mesenchymal stem cells in vitro and enhance im-munomodulatory functions. Stem Cells and Devel-opment. 23 (16), 1831-1843.

Nielsen, S., D. S., Nielsen, L. Lauritzen, M. Jakobsen, and K. F. Michaelsen. 2007. Impact of diet on the intes-tinal microbiota in 10-month-old infants. J. Pediatr. Gastroenterol. Nutr. 44:613-618.

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Nicholson, J. K., E. Holmes, J. Kinross, R. Burcelin, G. Gibson, W. Jia, and S. Pettersson, 2012. Host-gut microbiota metabolic interactions. Science. 336 (6086), 1262-1267.

Pluske, J. R. 2013. Feed- and feed additives-related as-pects of gut health and development in weanling pigs. J. Anim. Sci. Biotech. 4:1-7.

Pluske, J. R., D. L. Turpin, and J. Kim. 2018. Gastrointes-tinal tract (gut) health in the young pig. Anim. Nutr. 4:187-196.

Salcedo, J., S. A. Frese, D. A. Mills, and D. Barile. 2016. Characterization of porcine milk oligosaccharides during early lactation and their relation to the fecal microbiome. J. Dairy Sci. 99(10): 7733–7743.

Tao N., K. L. Ochonicky, J. B. German, S. M. Donovan, and C. B. Lebrilla. 2010. Structural determination and daily variations of porcine milk oligosaccha-rides. J. Agric. Food Chem. 58:4653–4659.

USDA. 2012. Part I: Baseline Reference of Swine Health and Environmental Management in the United States.

Yan H., and K. M. Ajuwon. 2017. Butyrate modifies in-testinal barrier function in IPEC-J2 cells through a selective upregulation of tight junction proteins and activation of the Akt signaling pathway. PLoS One. Jun 27;12(6):0179586.

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Glutamine and Transport StressAlan Duttlinger and Brian Richert

Department of Animal Sciences

Donald C. Lay Jr. and Jay S. Johnson USDA-ARS Livestock Behavior Research Unit

Purdue University West Lafayette, IN 47907


SummaryIn modern swine production, newly weaned pigs are often subjected to multiple stressors including weaning stress, transport stress, and thermal stress, and these have the potential to increase the incidence of animal disease, morbidity, and mortality, especially when they occur concomitantly. In order to promote stress recovery, prevent the onset of disease, and improve animal welfare, produc-ers often administer dietary antibiotics for 14 to 42 days after pigs enter nurseries or wean-to-finish facilities. However, due to increased consumer concern regarding the use of antibiotics in animal production and regulatory changes for antibiotic use, it has become increasingly important to de-velop antibiotic alternatives that can help pigs recover from stressful events as effectively as dietary antibiotics. It was determined in this study that replacing dietary antibiotics with 0.20% L-gluta-mine improved the productivity of weaned pigs at a similar level as dietary antibiotics throughout the nursery phase. However, the positive effects of dietary antibiotics and L-glutamine during the initial 14 days post-weaning on pig productivity were diminished as pigs entered the grow-finish phase, and no carcass characteristics were altered by nursery dietary treatments. Comparing sea-sons, pigs weaned in the spring had greater growth performance compared to pigs weaned in the summer resulting in heavier carcass weights and greater loin depth for spring weaned pigs. In con-clusion, 0.20% L-glutamine supplementation improved pig health and productivity after weaning and transport similarly to antibiotics; however, the positive growth effects of dietary antibiotics and L-glutamine provided the first 14 days post-weaning were diminished during the grow-finish phase.

Introduction Weaning is a complex stressor associated with so-cial, environmental, and metabolic stress in pigs (Lallés et al., 2004). In newly weaned pigs, stress is induced by separation from the sow, relocation and mixing piglet groups, and a radical change in diet that often reduces or eliminates feed intake in the first 48 hours post-weaning (Brooks et al., 2001). As a result, piglets un-dergo a variety of physiological and metabolic changes that can negatively impact welfare. Changes may result from elevated blood cortisol levels (Moeser et al., 2007), compromised feed intake (Maenz et al., 1994), altered intestinal morphology (Lallés et al., 2004), and dehydra-tion due to the switch from an all liquid (milk) to a solid diet. Unfortunately, in commercial production systems, weaning stress may be compounded by transport stress, which can induce significant weight loss with only 4 hours (h) of travel time (Hicks et al., 1998), and ambient

temperature likely plays a critical role in determining total stress load incurred by weaned piglets (Lambooy, 1988). Historically, swine producers used dietary antibiot-ics to help newly weaned pigs overcome the stress of weaning and associated stressors (Chiba, 2010). How-ever, due to several consumer and regulatory factors, it has become increasingly important to develop antibiot-ic alternatives that can help pigs recover from stressful events as effectively as dietary antibiotics. Previous re-search determined that inclusion of 0.20% L-glutamine in the diets of newly weaned and transported pigs could improve growth rate and well-being more effectively than dietary antibiotics (chlortetracycline + tiamulin; Johnson and Lay, 2017). However, this study was con-ducted under controlled conditions utilizing simulated transport and individual housing. Therefore, this study’s objectives were to evaluate the impact of replacing dietary antibiotics with 0.20% L-glutamine on swine

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growth performance, health status, welfare, and carcass characteristics of pigs in a production environment fol-lowing weaning and transport during different seasons (summer or spring). We hypothesized that withholding dietary antibiotics would negatively impact the overall well-being of piglets, and that diet supplementation with 0.20% L-glutamine would have a similar effect on piglet health and productivity as dietary antibiotics in a production environment.

Materials and Methods All procedures involving animal use were approved by the Purdue University Animal Care and Use Commit-tee and animal care and use standards were based upon the Guide for the Care and Use of Agricultural Animals in Research and Teaching (Federation of Animal Science Societies, 2010). Mixed sex crossbred pigs [n = 480; 5.62 ± 0.06 kg initial body weight (BW); Duroc x (Yorkshire x Landrace)] were weaned and transported at 18.4 ± 0.2 d of age in central Indiana during the summer of 2016 and the spring of 2017. One day prior to weaning and transport, all pigs were individually weighed, blocked by BW, and randomly allotted to pens, and pens of pigs were allotted to three dietary treatments with 10 pens per dietary treatment per season of weaning and trans-port. Initially, each pen contained eight pigs. Dietary treatments were antibiotics [A; chlortetracycline (441 ppm) + tiamulin (38.6 ppm)], no antibiotics (NA), or 0.20% L-glutamine (GLN; Ajinomoto North America, Inc., Raleigh, NC). The treatments were arranged in a 2 × 3 factorial with main effects of season (summer or spring) and diet (A, GLN, NA). On the day of weaning and transport, pigs were re-moved from sows and herded up a ramp into a goose-neck livestock trailer (2.35 × 7.32 m; Wilson Trailer Company, Sioux City, IA) providing 0.07 m2 per pig. The loading ramp to the trailer was 2.13 m in length provid-ing an 11.0⁰ incline. Two data loggers (Hobo®; data log-ger temperature/RH; Onset®; Bourne, MA) were evenly spaced within the trailer to measure ambient tempera-ture (TA) and relative humidity (RH) in 5 minute (m) intervals. During transport, the average trailer TA dur-ing the summer season was 29.4 ± 0.2°C and during the spring was 11.0 ± 0.2°C with trailer RH being 64.3 ± 0.8% and 63.1 ± 0.9%, respectively. Trailers were bedded with wood shavings and ventilation openings were adjusted based on the TA. Piglets were transported as a group in the trailer for approximately 12 h and 819 km without feed or water. Total transport time was determined by adding loading time, time spent in the trailer, unloading time, and the time it took to be sorted into their respective pens in the

nursery facility. The average time to wean and load the trailer was 55 m. The drivers were the same and followed the identical route for the summer and spring season. The transport route was approximately 50% two-lane roads and 50% four-lane roads. The route was approxi-mately 273 km in length and was completed three times during the transport phase for each season. The route took, on average 3 h 16 m to complete. The driver was changed and the truck was refueled after each time the 273 km route was completed. At the conclusion of the 12 h transport, piglets were unloaded from the trailer, individually weighed, and placed into pens. The average time to unload the trailer, weigh the pigs, and place into pens was 1 h 10 m.

Nursery Phase Following transport, pigs were placed in their as-signed pens and provided their respective dietary treat-ments for 14 d in two weekly phases (d 0 to 14 of the nursery phase). Following the dietary treatment period, all pigs were fed common antibiotic free diets from d 14 to the end of the nursery phase (d 34; Table 1). Diets were corn-soybean meal-based and formulated to meet or exceed nutrient requirements (NRC, 2012) in meal form in four phases during the nursery period (Table 1). Pig BW and feed consumption were recorded every 7 d during the nursery period to calculate ADG, ADFI, and G:F. On d 13 and 33, one pig per pen was harvested to collect plasma and gut tissue samples for biomarker as-says and intestinal histology. Therapeutic antibiotic administration was recorded for the duration of the trial (weaning to market). Pigs were treated when exhibiting clinical signs of illness. Treatment dose, product given, date given, pig and pen identification, and reason administered were recorded. Reason for therapeutic administration was then cat-egorized for post-hoc analysis. Categories were: enteric challenge (scours or loose watery stool), respiratory challenge (coughing, thumping, or labored breathing), lameness (carrying a limb or difficulty walking or swol-len joints), unthriftiness (BW loss, poor gain, loss of body condition, or rough hair coat), and all other treat-ments (Streptococcus suis, skin infection, and abscess). The nursery facility where the initial 34 d of the trial was conducted contained pens (1.22 m × 1.37 m) that provided initially approximately 0.21 m2 per pig. All pens contain one 5-hole dry self-feeder and a cup wa-terer to allow for ad libitum access to feed and water. The nursery barn has a shallow pit for manure storage and completely slatted plastic floors. The nursery room operated on mechanical ventilation using a 4-stage digi-tal controller (Airstream TC5-2V25A, Automated Pro-

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duction Systems, Assump-tion, IL, USA). During d 0 to 14 post-weaning, the nursery room average daily TA during the summer sea-son was 31.5 ± 1.82°C and during the spring was 30.6 ± 0.68°C. From d 14 to 34 the nursery TA was 28.7 ± 1.14°C and 26.0 ± 0.84°C for the summer and spring seasons, respectively.

Grow-Finish Phase On day 34, all pigs were moved to the grow-finish facility for the remainder of the trial and pen in-tegrity was maintained. Common antibiotic free diets were corn-soybean meal-DDGS-based diets provided in meal form to meet or exceed nutrient re-quirements (NRC, 2012) in six phases during the grow-finish period (Table 2). Pigs and feeders were weighed every 21 d during the grow-finish period to determine ADG, ADFI, and G:F. The grow-finish facility contained pens (1.68 m × 4.27 m) that provided ap-proximately 1.19 m2 per pig. All pens contained one 2-hole dry self-feeder and a nipple waterer to al-low for ad libitum access to feed and water. The grow-finish barn had a shallow pit for manure storage and completely slatted concrete floors. The barn was mechanically venti-lated. During d 0 to 62 of the grow-finish phase, the

Table 1. Composition of Nursery Diets, as fed.

ItemPhase 11

Phase 22

Phase 33 Phase 44 A5 GLN6 NA7 A GLN NAIngredient, % Corn 30.81 31.18 31.38 37.52 37.89 38.09 51.63 57.38SBM, 48% CP 13.95 13.95 13.95 18.00 18.00 18.00 25.65 30.70DDGS, 7% oil --- --- --- --- --- --- --- 5.00Soybean Oil 5.00 5.00 5.00 5.00 5.00 5.00 3.00 ---Choice White Grease --- --- --- --- --- --- --- 3.00Limestone 0.79 0.79 0.79 0.74 0.74 0.74 0.86 1.33Monocal. Phos. 0.40 0.40 0.40 0.49 0.49 0.49 0.49 0.74Vitamin Premix8 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.25Trace Mineral Premix9 0.125 0.125 0.125 0.125 0.125 0.125 0.125 0.125Selenium Premix10 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05Phytase11 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10Salt 0.25 0.25 0.25 0.25 0.25 0.25 0.30 0.35Plasma Protein 6.50 6.50 6.50 2.50 2.50 2.50 --- ---Spray Dried Blood Meal 1.50 1.50 1.50 1.50 1.50 1.50 --- ---Soy Concentrate 4.00 4.00 4.00 3.00 3.00 3.00 2.50 ---Select Menhaden Fish Meal

5.00 5.00 5.00 4.00 4.00 4.00 4.00 ---

Dried Whey 25.00 25.00 25.00 25.00 25.00 25.00 10.00 ---Lactose 5.00 5.00 5.00 --- --- --- --- ---L-Lysine-HCL 0.07 0.07 0.07 0.2 0.2 0.2 0.28 0.40DL-Methionine 0.22 0.22 0.22 0.23 0.23 0.23 0.18 0.17L-Threonine 0.04 0.04 0.04 0.09 0.09 0.09 0.12 0.14L-Tryptophan --- --- --- 0.01 0.01 0.01 0.01 ---Zinc Oxide 0.375 0.375 0.375 0.375 0.375 0.375 0.375 ---Copper Sulfate --- --- --- --- --- --- --- 0.10Aureomycin 5012 0.40 --- --- 0.40 --- --- --- ---Denagard 1013 0.18 --- --- 0.18 --- --- --- ---L-Glutamine14 --- 0.20 --- --- 0.20 --- --- ---Banminth 4815 --- --- --- --- --- --- --- 0.10Clarifly, 0.67%16 --- --- --- --- --- --- 0.08 0.07

Calculated chemical compositionME, kcal/kg 3536 3536 3536 3510 3510 3510 3418 3396Fat, % 7.27 7.27 7.27 7.36 7.36 7.36 5.73 5.86CP, % 24.6 24.6 24.6 22.9 22.9 22.9 22.3 21.3SID Lysine, % 1.55 1.55 1.55 1.45 1.45 1.45 1.35 1.25Ca, % 0.90 0.90 0.90 0.85 0.85 0.85 0.80 0.75Total P, % 0.75 0.75 0.75 0.71 0.71 0.71 0.64 0.57Avail. P, % 0.60 0.60 0.60 0.55 0.55 0.55 0.45 0.36

1 Fed d 0 to 7 post-weaning and transport.2 Fed d 7 to 14 post-weaning and transport.3 Fed d 14 to 21 post-weaning and transport.4 Fed d 21 to 34 post-weaning and transport.5 Pigs provided dietary antibiotics [chlortetracycline (441 ppm) + tiamulin (38.6 ppm)].6 Pigs provided 0.20% L-glutamine 7 Pigs provided no dietary antibiotics8 Provided per kilogram of the diet: vitamin A, 6,614 IU; vitamin D3, 661 IU; vitamin E, 44 IU; vitamin

K, 2.2 mg; riboflavin, 8.8 mg; pantothenic acid, 22 mg; niacin, 33 mg; vitamin B12, 0.039 mg.9 Provided per kilogram of the diet available: iron, 121.3 mg; zinc, 121.3 mg; manganese, 15.0 mg;

copper, 11.3 mg; iodine, 0.46 mg.10 Provided 0.3 ppm selenium11 Provided 600 FTU per kg of the diet12 Aureomycin (Zoetis, Parsippany, NJ) provided 441 ppm chlortetracycline in the diet.13 Denagard (Elanco Animal Health, Greenfield, IN) provided 38.6 ppm tiamulin in the diet.14 Ajinomoto North America, Inc., Raleigh, NC15 Banminth (Phibro Animal Health Corporation, Teaneck, NJ) provided 106 ppm pyrantel tartrate in

the diet.16 Clarifly (Central Life Sciences, Schaumburg, IL) provided 5.4 ppm (Phase 3) and 4.7 ppm (Phase 4)

diflubenzuron in the diet.

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room average daily TA for the summer weaned pigs was 22.4 ± 1.14°C and for the spring weaned pigs was 25.5 ± 2.64°C. From d 62 to 125 the TA was 19.9 ± 0.83°C and 25.7 ± 2.48°C for the pigs weaned in the summer and spring sea-sons, respectively.

Marketing At the end of the 159 d experiment, pigs from each pen were tattooed with pen number and shipped approximately 48 km to Indiana Pack-ers Corporation (Delphi, IN). Pigs were slaughtered under commercial condi-tions with carbon dioxide stunning. Standard carcass criteria of loin and backfat depth, hot carcass weight (HCW), fat-free lean in-dex, and yield were col-lected. Fat depth and loin depth were measured with an optical probe (Fat-O-Meater, SFK Technology A/S, Herlev, Denmark) inserted between the third and fourth rib from the last rib (counting from the posterior of the carcass) and 7 cm from the dorsal midline of the hot carcass. Lean percentage was cal-culated according to the Indiana Packers Corpora-tion (2015) formula and the fat-free lean percent-age was calculated ac-cording to Schinckel et al. (2010) procedures.

Statistics Data were analyzed as a randomized complete block design using the PROC MIXED procedure in SAS 9.4 (SAS Institute INC., Cary, NC), with pen as the experi-mental unit. Main effects of season and diet and their

Table 2. Composition of grow-finish diets, as fed.

Item Phase 11 Phase 22 Phase 33 Phase 44 Phase 55 Phase 66

Ingredient, % Corn 61.47 64.65 66.40 71.10 82.38 68.67SBM, 48% CP 23.20 16.15 9.75 5.25 4.25 15.10DDGS, 7% Fat 10.00 15.00 20.00 20.00 10.00 10.00Choice White Grease 2.00 1.00 1.00 1.00 1.00 3.00Limestone 1.37 1.35 1.39 1.32 1.16 1.26Monocal. Phos. 0.47 0.32 0.05 0.00 0.10 0.27Vitamin Premix 0.1507 0.1507 0.1258 0.1209 0.10010 0.1507

Trace Mineral Premix 0.1011 0.0912 0.0813 0.0714 0.0515 0.1011

Selenium Premix 0.0516 0.0516 0.0516 0.0516 0.02517 0.0516

Phytase18 0.10 0.10 0.10 0.10 0.10 0.10Salt 0.35 0.35 0.30 0.30 0.25 0.30L-Lysine-HCL 0.42 0.46 0.48 0.46 0.37 0.42DL-Methionine 0.11 0.08 0.05 0.01 0.00 0.10L-Threonine 0.130 0.130 0.120 0.105 0.095 0.160L-Tryptophan 0.010 0.030 0.035 0.040 0.030 0.030Paylean 2.2519 --- --- --- --- --- 0.15Availa Zn 12020 --- --- --- --- --- 0.042Clarifly, 0.67%21 0.07 0.09 0.07 0.08 0.09 0.10

Calculated chemical compositionME, kcal/kg 3373 3337 3351 3359 3371 3438Fat, % 5.29 4.69 5.06 5.15 4.73 6.40CP, % 19.34 17.59 15.99 14.18 11.90 16.01SID Lysine, % 1.10 0.98 0.85 0.73 0.60 0.90Ca, % 0.70 0.65 0.60 0.55 0.50 0.60Total P, % 0.50 0.47 0.41 0.38 0.35 0.42Avail. P, % 0.32 0.30 0.26 0.24 0.20 0.26

1 Fed d 0 to 21 of the grow-finish phase.2 Fed d 21 to 42 of the grow-finish phase.3 Fed d 42 to 62 of the grow-finish phase.4 Fed d 62 to 83 of the grow-finish phase.5 Fed d 83 to 104 of the grow-finish phase.6 Fed d 104 to 125 of the grow-finish phase.7 Provided per kilogram of the diet: vitamin A, 3,968 IU; vitamin D3, 397 IU; vitamin E, 26.5 IU; vitamin

K, 1.3 mg; riboflavin, 5.3 mg; pantothenic acid, 13.2 mg; niacin, 19.8 mg; vitamin B12, 0.023 mg.8 Provided per kilogram of the diet: vitamin A, 3,307 IU; vitamin D3, 331 IU; vitamin E, 22.1 IU; vita-

min K, 1.1 mg; riboflavin, 4.4 mg; pantothenic acid, 11 mg; niacin, 16.5 mg; vitamin B12, 0.020 mg.9 Provided per kilogram of the diet: vitamin A, 3,174 IU; vitamin D3, 317 IU; vitamin E, 21.2 IU; vitamin

K, 1.0 mg; riboflavin, 4.2 mg; pantothenic acid, 10.6 mg; niacin, 15.8 mg; vitamin B12, 0.019 mg.10 Provided per kilogram of the diet: vitamin A, 2,647 IU; vitamin D3, 265 IU; vitamin E, 17.7 IU; vitamin

K, 0.9 mg; riboflavin, 3.5 mg; pantothenic acid, 8.8 mg; niacin, 13.2 mg; vitamin B12, 0.016 mg.11 Available mineral provided per kilogram of the diet: iron, 97 mg; zinc, 97 mg; manganese, 12 mg;

copper, 9 mg; iodine, 0.37 mg.12 Available mineral provided per kilogram of the diet: iron, 87 mg; zinc, 87 mg; manganese, 10.8 mg;

copper, 8.1 mg; iodine, 0.33 mg.13 Available mineral provided per kilogram of the diet: iron, 77.6 mg; zinc, 77.6 mg; manganese, 9.6

mg; copper, 7.2 mg; iodine, 0.29 mg.14 Available mineral provided per kilogram of the diet: iron, 68 mg; zinc, 68 mg; manganese, 8.4 mg;

copper, 6.3 mg; iodine, 0.26 mg.15 Available mineral provided per kilogram of the diet: iron, 48.5 mg; zinc, 48.5 mg; manganese, 6 mg;

copper, 4.5 mg; iodine, 0.18 mg.16 Provided 0.3 ppm selenium17 Provided 0.15 ppm selenium18 Provided 600 FTU per kg of the diet19 Paylean (Elanco Animal Health, Greenfield, IN) provided 7.5 ppm ractopamine HCl in the diet20 Zinpro Corporation, Eden Prairie, MN21 Clarifly (Central Life Sciences, Schaumburg, IL) provided 4.7, 6.0, 5.4, 6.7 ppm diflubenzuron in the

diet, respectively.

interactions were tested. The assumptions of normality of error, homogeneity of variance, and linearity were confirmed post-hoc. All therapeutic treatment rate data were log-transformed to meet assumptions of normal-

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ity; however, all log-transformed data are presented as arithmetic means for ease of interpretation. All non-transformed data are presented as LS means. Statistical significance was defined as P ≤ 0.05 and a tendency was defined as 0.05 < P ≤ 0.10.

ResultsNursery Phase When comparing the main effect of dietary treat-ment, ADG was greater overall (P = 0.01; 14.9%) from d 0 to 14 of the nursery period in A and GLN fed pigs compared to NA pigs, but no ADG differences were de-tected between A and GLN pigs (Table 3). An increase in ADFI (P = 0.04) was detected from d 0 to 14 of the nursery phase for A compared to NA pigs, but no dif-ferences were observed between GLN versus A and NA pigs. Feed efficiency (G:F) was greater (P = 0.01; 7.7%) from d 0 to 14 of the nursery phase for A compared to NA and GLN pigs, but no differences were observed between NA and GLN pigs. Day 14 BW was greater overall (P = 0.01) for A (8.65 kg) and GLN (8.50 kg) pigs compared to NA (8.19 kg) pigs; however, no differences were detected between A and GLN pigs (Table 3). Overall, from d 0 to 34 of the nursery period, ADG (P = 0.01; 7.9%), ADFI (P = 0.09), G:F (P = 0.01; 4.3%), and final BW (P = 0.04) were greater for pigs fed A compared to NA fed pigs, with GLN fed pigs being intermediate and not different from A and NA pigs (Table 3). No oth-er dietary treatment growth performance differences (P > 0.05) were detected during the nursery phase. During the spring season, pigs tended to have re-duced ADFI (P = 0.08; 5.1%) compared to the summer from d 0 to 14 of the nursery phase (Table 3). However, from d 14 to 34 of the nursery phase, ADG tended to be increased (P = 0.09) and G:F was increased (P = 0.01) during the spring compared to the summer (3.7 and 7.4%, respectively; Table 3). Overall, from d 0 to 34 of the nursery period, G:F was reduced (P = 0.04; 4.1%) during the summer compared to the spring (Table 3). No other seasonal effects were observed during the nursery pe-riod (P > 0.05). A diet x season interaction was detected (P = 0.04) from d 14 to 34 of the nursery phase where G:F was greater in the spring in NA (0.69 ± 0.01) and GLN (0.68 ± 0.01) pigs compared to NA pigs (0.61 ± 0.01) during the summer. However, no differences were observed between A pigs (0.66 ± 0.01) during the spring and A (0.64 ± 0.01) and GLN (0.63 ± 0.01) pigs during the sum-mer (data not presented). No other diet x season inter-actions were detected (P < 0.05).

On d13, plasma TNF-α was reduced (P=0.02) in A (36.7±6.9 pg/mL) and GLN pigs (40.9±6.9 pg/mL) versus NA pigs (63.2±6.9 pg/mL). On d33, villus height:crypt depth tended (P=0.09) to be greater in A (2.71±0.09) and GLN (2.72±0.09) compared to NA (2.54±0.09) fed pigs.

Grow-Finish Phase There were no carry-over effects of nursery dietary treatments observed (P > 0.17) during the grow-finish period (Table 4). From d 0 to 62 of the grow-finish phase, G:F was reduced (P = 0.01; 4.3%) for the pigs weaned during the summer compared to the spring (Table 4). Average daily gain, ADFI, and G:F were reduced (P = 0.01; 14.6, 4.4, and 12.1%, respectively) for the summer weaned pigs from d 62 to 125 of the grow-finish phase compared to the spring pigs (Table 4). Overall, from d 0 to 125 of the grow-finish period, ADG, G:F, and final BW were reduced (P = 0.01; 9.2%, 5.1%, and 9.8 kg, re-spectively) in the summer weaned pigs compared to the spring (Table 4). No other growth performance differ-ences were observed (P > 0.05) during the grow-finish period with any comparison (Table 4).

Treatment rates From d 0 to 14, there was a tendency (P = 0.07) for a diet by season interaction for enteric treatments with pigs fed A having the lowest and similar percentage of pigs treated in the summer (3.13%) and spring (3.13%), however pigs fed GLN had a similar low percentage of pigs treated during the summer (3.75%) but the greatest percentage in the spring (8.19%) while the pigs fed NA diets had the greatest (6.88%) enteric therapeutic treat-ments during the summer and a reduced intermediate level of therapies (4.55%) in the spring (Table 5). Pigs treated for other reasons were greater (P ≤ 0.02) from d 0 to 14 during the spring (1.08%) compared to the sum-mer (0.00%), regardless of dietary treatment (Table 5). A diet x season effect was detected (P = 0.04) where pigs treated for lameness from d 14 to 34 was greater in the spring for GLN pigs (2.12%) compared to all other treatments. However, no differences were observed be-tween A (0.56%) and NA (0.00%) pigs during the spring, and A (0.48%), GLN (0.00%), and NA (0.00%) pigs dur-ing the summer (Table 5). During d 62 to 125 of the grow-finish phase, enteric disease treatments tended (P < 0.08) to be reduced by A (0.00%) treatment and highest for the GLN (1.17%) treatment with NA (0.34%) pigs being intermediate (Ta-ble 6). From d 62 to 125, treatment for unthriftiness was reduced (P = 0.01) in GLN (0.00%) and NA pigs (0.00%) compared to A pigs (1.11%).

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Table 3. Effect of dietary treatment and season on nursery pig growth performance.1

Parameter

Main Effects

SEPSeason

Diet

Summer2 Spring3 A4 GLN5 NA6 D7 S8 D x SDay 0 to 14

Initial BW, kg 5.64 5.51 5.58 5.59 5.57 0.29 0.99 0.70 0.99ADG, g 210.1 206.1 224.2a 210.8a 189.2b 10.19 0.01 0.56 0.82ADFI, g 274.6 260.6 277.1a 272.1ab 253.6b 13.21 0.04 0.08 0.92G:F 0.80 0.80 0.84a 0.79b 0.77b 0.01 0.01 0.91 0.17Day 14 BW, kg 8.44 8.46 8.65a 8.50a 8.19b 0.52 0.01 0.83 0.97

Day 14 to 34ADG, g 439.0 455.7 458.0 447.4 436.7 12.05 0.21 0.09 0.43ADFI, g 693.5 674.7 702.3 680.4 669.6 22.81 0.16 0.19 0.63G:F 0.63 0.68 0.65 0.66 0.65 0.01 0.78 0.01 0.04

Day 0 to 34ADG, g 347.4 355.9 364.5a 352.7ab 337.7b 10.18 0.01 0.23 0.58ADFI, g 525.9 509.0 532.2x 517.1xy 503.2y 17.43 0.09 0.12 0.77G:F 0.70 0.73 0.73a 0.71ab 0.70b 0.01 0.03 0.01 0.07Final BW, kg 17.20 17.62 17.78a 17.49ab 16.96b 0.74 0.04 0.11 0.69

1 A total of 10 pens were used per dietary treatment per season.2 Pigs weaned and transported for 12 h during July 2016.3 Pigs weaned and transported for 12 h during April 2017.4 Pigs provided dietary antibiotics [chlortetracycline (441 ppm) + tiamulin (38.6 ppm)] for 14 d post-wean-

ing and transport and then fed common antibiotic free diets.5 Pigs provided 0.20% L-glutamine for 14 d post-weaning and transport and then fed common antibiotic

free diets.6 Pigs provided no dietary antibiotics for 14 d post-weaning and transport and then fed common antibiotic

free diets.7 Dietary treatment8 Seasona,b Letters indicate significant differences (P ≤ 0.05) within a row and main effect.x,y Letters indicate tendencies (0.05 < P ≤ 0.10) within a row and main effect.

Table 4. Effect of dietary treatment and season on grow-finish pig growth performance.1

Parameter

Main Effects

SEPSeason

Diet

Summer2 Spring3 A4 GLN5 NA6 D7 S8 D x SDay 0 to 62

ADG, kg 0.76 0.77 0.78 0.76 0.76 0.01 0.32 0.37 0.62ADFI, kg 1.79 1.75 1.80 1.76 1.75 0.03 0.40 0.14 0.88G:F 0.44 0.46 0.45 0.46 0.45 0.01 0.80 0.01 0.36Day 62 BW, kg 64.72 65.50 65.99 65.02 64.31 0.96 0.22 0.32 0.76

Day 62 to 125ADG, kg 0.82 0.96 0.88 0.89 0.90 0.02 0.41 0.01 0.36ADFI, kg 2.83 2.96 2.87 2.91 2.90 0.05 0.72 0.01 0.42G:F 0.29 0.33 0.30 0.31 0.31 0.01 0.17 0.01 0.62

Day 0 to 125ADG, kg 0.79 0.87 0.83 0.83 0.83 0.01 0.95 0.01 0.58ADFI, kg 2.31 2.35 2.33 2.33 2.32 0.03 0.97 0.21 0.60G:F 0.37 0.39 0.38 0.38 0.38 0.01 0.54 0.01 0.56Final BW, kg 117.37 127.19 122.77 121.73 122.34 1.23 0.83 0.01 0.64

1 A total of 10 pens were used per dietary treatment per season.2 Pigs weaned and transported for 12 h during July 2016.3 Pigs weaned and transported for 12 h during April 2017.4 Pigs provided dietary antibiotics [chlortetracycline (441 ppm) + tiamulin (38.6 ppm)] for 14 d post-wean-

ing and transport and then fed common antibiotic free diets.5 Pigs provided 0.20% L-glutamine for 14 d post-weaning and transport and then fed common antibiotic

free diets.6 Pigs provided no dietary antibiotics for 14 d post-weaning and transport and then fed common antibiotic

free diets.7 Dietary treatment8 Season

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Table 5. Effect of dietary treatment and season on therapeutic antibiotic treatment rate during the nursery period.1

Parameter

Main Effects

SEPSummer2

Spring3

A4 GLN5 NA6 A GLN NA D7 S8 D x SDay 0 to 14

Enteric9 3.13y 3.75y 6.88xy 3.13y 8.19x 4.55xy 2.31 0.31 0.38 0.07Lame10 1.88 1.88 1.25 0.63 1.39 0.63 1.02 0.73 0.27 0.89Unthrifty11 1.88 1.25 1.25 0.00 0.69 1.25 1.02 0.92 0.22 0.48Respiratory12 --- --- --- --- --- --- --- --- --- ---Other13 0.00 0.00 0.00 1.25 0.69 1.25 0.86 0.86 0.02 0.86

Day 14 to 34Enteric 0.00 0.95 0.48 0.00 0.00 0.56 0.66 0.36 0.33 0.37Lame 0.48b 0.00b 0.00b 0.56b 2.12a 0.00b 1.00 0.08 0.06 0.04Unthrifty 0.00 0.00 1.03 1.03 1.15 1.03 0.80 0.64 0.14 0.57Respiratory 0.00 0.00 0.48 0.00 0.53 0.00 0.53 0.58 0.94 0.20Other 0.00 0.00 0.00 0.00 0.53 0.00 0.53 0.31 0.27 0.31

1 A total of 10 pens were used per dietary treatment per season.2 Pigs weaned and transported for 12 h during July 2016.3 Pigs weaned and transported for 12 h during April 2017.4 Pigs provided dietary antibiotics [chlortetracycline (441 ppm) + tiamulin (38.6 ppm)] for 14 d post-weaning and

transport and then fed common antibiotic free diets.5 Pigs provided 0.20% L-glutamine for 14 d post-weaning and transport and then fed common antibiotic free diets.6 Pigs provided no dietary antibiotics for 14 d post-weaning and transport and then fed common antibiotic free

diets.7 Dietary treatment8 Season9 Percent of pigs within pen treated with therapeutic antibiotics for enteric challenge.10 Percent of pigs within pen treated with therapeutic antibiotics for lameness.11 Percent of pigs within pen treated with therapeutic antibiotics for un-thriftiness.12 Percent of pigs within pen treated with therapeutic antibiotics for respiratory challenge.13 Percent of pigs within pen treated with therapeutic antibiotics for all other conditions.a,b,c Letters indicate significant differences (P ≤ 0.05) within a row.x,y Letters indicate tendencies (0.05 < P ≤ 0.10) within a row.

Table 6. Effect of season and dietary treatment on therapeutic antibiotic treatment rate for enteric challeng-es during the grow-finish period.1

ParameterSummer2

Spring3

SEP

A4 GLN5 NA6 A GLN NA D7 S8 D x SDay 0 to 62

Enteric9 0.56 0.00 0.00 0.56 0.67 0.67 0.67 0.81 0.31 0.77Lame10 0.56 0.56 1.89 0.00 0.00 0.00 1.06 0.41 0.02 0.41Unthrifty11 1.78 0.00 0.67 0.00 0.00 0.56 0.99 0.24 0.17 0.16Respiratory12 11.11 10.78 8.00 0.56 1.11 2.22 2.92 0.60 <0.01 0.77Other13 1.11 2.33 1.33 0.00 0.00 0.56 1.11 0.69 0.02 0.45

Day 62 to 125Enteric 0.00 0.67 0.00 0.00 1.67 0.67 0.93 0.08 0.19 0.58Lame 2.22 0.67 0.67 0.00 0.00 0.00 1.05 0.21 0.01 0.21Unthrifty 0.56 0.00 0.00 1.67 0.00 0.00 0.93 0.01 0.28 0.30Respiratory 10.33 8.22 12.17 7.33 7.39 6.78 4.20 0.81 0.49 0.86Other 0.56 0.00 0.00 0.00 0.00 0.00 0.56 0.37 0.32 0.37

1 A total of 10 pens were used per dietary treatment per season.2 Pigs weaned and transported for 12 h during July 2016.3 Pigs weaned and transported for 12 h during April 2017.4 Pigs provided dietary antibiotics [chlortetracycline (441 ppm) + tiamulin (38.6 ppm)] for 14 d post-weaning and

transport and then fed common antibiotic free diets.5 Pigs provided 0.20% L-glutamine for 14 d post-weaning and transport and then fed common antibiotic free diets.6 Pigs provided no dietary antibiotics for 14 d post-weaning and transport and then fed common antibiotic free

diets.7 Dietary treatment8 Season9 Percent of pigs within pen treated with therapeutic antibiotics for enteric challenge.10 Percent of pigs within pen treated with therapeutic antibiotics for lameness.11 Percent of pigs within pen treated with therapeutic antibiotics for un-thriftiness.12 Percent of pigs within pen treated with therapeutic antibiotics for respiratory challenge.13 Percent of pigs within pen treated with therapeutic antibiotics for all other conditions.

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Pigs treated for lameness were greater (P < 0.02) from d 0 to 62 and d 62 to 125 during the summer (1.00%, 1.19%, respectively) compared to the spring (0.00%, 0.00%, respectively), regardless of dietary treatment (Ta-ble 6). Treatment for respiratory challenges were great-er (P < 0.01) from d 0 to 62 during the summer (9.96%) compared to the spring (1.30%). Pigs treated for other challenges were also greater (P < 0.02) during the sum-mer compared to the spring from d 0 to 62.

Carcass Characteristics No dietary treatment effects were observed (P > 0.60) on carcass characteristics (Table 7). Hot carcass weight and loin depth were increased (P < 0.01; 5.4% and 5.5%, respectively) and carcass yield was reduced (P < 0.01; 2.0%) for pigs weaned during the spring compared to the summer (Table 7). When HCW was used as a co-variate, loin depth and lean percentage were increased (P = 0.01; 4.0% and 1.1%, respectively) and carcass yield was reduced (P = 0.01; 2.3%) for the spring weaned pigs compared to the summer (Table 7). Fat-free lean per-centage for the spring weaned pigs tended to be greater (P = 0.07; 1.3%) compared to the summer when HCW was included as a covariate (Table 7).

Discussion The need to wean and transport pigs is necessary to reduce the risk of infectious disease through multi-site production (Harris, 2000). However, the resultant stress response can reduce growth performance and welfare in newly weaned pigs (Campbell et al., 2013), especially in the absence of dietary antibiotics (Heo et al., 2013). Despite this, the use of in-feed antibiotics has been reduced in swine production due to consumer preference, legislative action, and concerns about anti-biotic resistance, putting the welfare and productivity of newly weaned and transported pigs at risk and necessi-tating the development of effective alternatives. Recent work has described improved welfare and productivity in piglets provided GLN compared to A and NA follow-ing weaning and simulated transport (Johnson and Lay, 2017). In accordance with the aforementioned study, piglets provided GLN after weaning and transport in the present study had improved growth performance compared to NA pigs during the 14 d dietary treatment period, regardless of season of transport. Additionally, no growth performance differences were detected be-tween GLN and A pigs in our current study. Although reasons for this discrepancy are currently unknown, it may be due to differences in study design since the

Table 7. Effect of season and dietary treatment on carcass characteristics.1

Parameter

Main Effects

SEPSeason Diet

Summer2 Spring3 A4 GLN5 NA6 D7 S8 D x SNo HCW9 covariate

HCW, kg 92.42 97.44 95.32 95.54 93.93 1.32 0.60 <0.01 0.70Loin depth, mm 64.0 67.4 65.8 65.9 65.5 0.72 0.93 <0.01 0.60Backfat, mm 21.4 22.1 21.7 21.6 21.7 0.59 0.99 0.31 0.40Yield, % 77.2 75. 7 76.6 76.4 76.4 0.19 0.68 <0.01 0.46Lean, %10 54.42 54.61 54.51 54.55 54.47 0.25 0.97 0.53 0.54Fat-free lean, %11 48.69 48.79 48.74 48.79 48.69 0.30 0.97 0.76 0.50

HCW covariateLoin depth, mm 64.4 67.00 65.7 65.7 65.7 0.69 0.99 0.01 0.66Backfat, mm 22.0 21.4 21.6 21.5 22.0 0.49 0.75 0.33 0.57Yield, % 77.3 75.5 76.5 76.3 76.4 0.17 0.69 0.01 0.63Lean, % 54.20 54.82 54.54 54.60 54.38 0.23 0.78 0.04 0.71Fat-free lean, % 48.41 49.06 48.77 48.85 48.58 0.27 0.77 0.07 0.68

1 A total of 10 pens were used per dietary treatment per season.2 Pigs weaned and transported for 12 h during July 2016.3 Pigs weaned and transported for 12 h during April 2017.4 Pigs provided dietary antibiotics [chlortetracycline (441 ppm) + tiamulin (38.6 ppm)] for 14 d post-weaning

and transport and then fed common antibiotic free diets.5 Pigs provided 0.20% L-glutamine for 14 d post-weaning and transport and then fed common antibiotic free

diets.6 Pigs provided no dietary antibiotics for 14 d post-weaning and transport and then fed common antibiotic

free diets.7 Dietary treatment8 Season9 Hot carcass weight10 Equation used: 54.672154 - (0.412525 × backfat, mm) - (0.002982 × hot carcass weight, kg × 2.20462) +

(0.1433242 × loin depth, mm) (Indiana Packers Corporation, 2015)11 Equation used: 51.2 - (0.510 × backfat, mm) + (0.131 × loin depth, mm) (Schinckel et al., 2010)

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transport procedure was simulated and piglets were individually housed in the previous study (Johnson and Lay, 2017). While the mechanism(s) of action for improved growth performance has yet to be fully dis-cerned, GLN can serve as energy source for enterocytes thus reducing jejunal atrophy and intestinal epithelial damage (Wu et al., 1996; Yi et al., 2005). Therefore, it is possible that piglets provided supplemental GLN had improved intestinal barrier function leading to greater pathogen resistance, reduced translocation of bacte-ria (Peng, 2004) and subsequently an improvement in growth performance (Jiang et al., 2009; Johnson and Lay, 2017). This is partially supported by the greater vil-lus height:crypt depth ratios of pigs fed GLN and A in our study. Additionally, we observed at the end of the di-etary feeding period (d13), plasma TNF-α was reduced 38.6% for pigs fed GLN and A, further supporting these dietary effects on improving intestinal health and func-tion by feeding GLN and A. These advantages observed in early nursery growth performance may suggest that GLN supplementation could serve as an alternative to dietary antibiotics in production systems. Although growth performance was improved in GLN and A pigs during the dietary treatment period and the advantage was maintained for the overall nurs-ery period, no differences were detected when com-pared to NA pigs from d 34 to market when all pigs were fed common antibiotic free diets during the grow-finish period. However, these results were somewhat expect-ed as previous research has described a loss of growth performance differences once dietary antibiotic treat-ments ceased (Skinner et al., 2014). This may be due to variability differences that diminished the growth rate advantages as the studies progressed or the per-formance advantages of feeding dietary treatments are limited only to the period when fed. Therefore, it could be suggested that feeding GLN to pigs for a longer dura-tion could have extended the growth benefits; however, further work would be needed to confirm this hypothe-sis and any increase in growth performance would need to be balanced with the cost of including GLN in diets. Therapeutic injectable antibiotics are one of many options currently available to aid in the control of pathogens and disease in addition to good biosecurity practices, vaccinations, and dietary antibiotics. In the present study, pigs receiving A had fewer therapeutic antibiotic treatments for enteric challenges compared to GLN pigs during the spring from d 0 to 14 post-weaning, but no differences were detected during the summer. While this may indicate that dietary antibiotic treatments were more effective at reducing pathogen

impact compared to GLN, the lack of overall dietary treatment differences may suggest that the season of weaning and transport influences the impact of GLN on therapeutic treatments. Regardless, the increase in ther-apeutic treatments did not appear to coincide with a re-duction in growth performance and this may be due to differences in the mode of action between A and GLN treatments whereby dietary antibiotics reduce patho-gen colonization (Pluske et al., 2002) and GLN fed pigs may improve gut barrier function (Wang et al., 2015). In the present study, no dietary treatment carcass trait differences were detected, confirming previous reports that providing dietary antibiotics for a limited period in the nursery phase would have no impact on meat quality (Skinner et al., 2014). While the effects of providing GLN on carcass characteristics in pigs are un-known, previous reports in broilers reported that GLN supplementation during heat stress improved meat yield (Dai et al., 2011). However, because broilers were provided GLN until harvest in the aforementioned study and pigs in the present study were only provided GLN for 14 d, it is likely that the lack of carcass trait dif-ferences are related to the duration of dietary inclusion. Despite the lack of dietary treatment differences on carcass characteristics, pigs weaned in the spring had greater HCW and loin depth and increased lean percentage and fat-free lean percentage when HCW was used as a covariate compared to summer weaned pigs. While the mechanism(s) for the improvement in carcass characteristics are unknown, we speculate that health status may have impacted the carcass differences observed in the current study due to the differences in therapeutic antibiotic treatment rate between seasons. This response appears to be consistent with previous work by Holck et al., (1998) and Williams et al., (1997) who reported improved carcass characteristics when pigs were reared under higher health status. This sug-gests that poorer health status may have decreased growth rate and subsequently reduced lean tissue accre-tion rate. This potential advantage in health status dur-ing the spring may have allowed the pigs to grow and deposit lean tissue at a rate closer to their genetic poten-tial because previous studies determined that when pigs were exposed to chronic immune system activation in a health compromised environment, cytokine concen-tration was elevated (Williams et al., 1997) thereby sup-pressing lean growth. Less environmental pathogens as indicated by reduced therapeutic antibiotic use could have decreased immune system and cytokine activation thus allowing the potential for increased muscle accre-tion rate.

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Conclusion Weaning and transport is stressful to pigs and an-tibiotics have been routinely used to help young pigs overcome these challenges. Despite the advantages in growth performance and productivity found from the use of dietary antibiotics, alternatives to antibiotics are needed. We determined that L-glutamine supple-mented at 0.20% improved pig health and productivity after weaning and transport similarly to antibiotics dur-ing the nursery phase; however, the positive effects of early nursery dietary antibiotics and L-glutamine were diminished during the grow-finish phase. However, pigs not provided dietary antibiotics had decreased growth rate during the nursery phase. Future work should ad-dress if the benefits of feeding dietary antibiotics and L-glutamine are additive and to better understand the mechanism involved for L-glutamine supplementation to improve pig growth performance following weaning and transport stressors.

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A. Campbell. 2001. Liquid feeding for the young pig-let. In The Weaner Pig: Nutrition and Management, pp. 153–178 [MA Varley and J Wiseman, eds]. Wallingford: CABI Publishing.

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Johnson, J.S., and D.C. Lay Jr. 2017. Evaluating the be-havior, growth performance, immune parameters, and intestinal morphology of weaned piglets after simulated transport and heat stress when antibiot-ics are eliminated from the diet or replaced with L-glutamine. J. Anim. Sci. 95: 91-102.

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Schinckel, A. P., J. R. Wagner, J. C. Forrest, and M. E. Ein-stein. 2010. Evaluation of the prediction of alterna-tive measures of pork carcass composition by three optical probes. J. Anim. Sci. 2010. 88:767–794.

Skinner, L. D., C. L. Levesque, D. Wey, M. Rudar, J. Zhu, S. Hooda, and C. F. M. de Lange. 2014. Impact of nursery feeding program on subsequent growth performance, carcass quality, meat quality, and physical and chemical body composition of grow-ing-finishing pigs. J. Anim. Sci. 92:1044-1054.

Wang, H., C. Zhang, G. Wu, Y. Sun, B. Wang, B. He. Z. Dai, and Z. Wu. 2015. Glutamine enhances tight junction protein expression and modulates cortico-tropin-releasing factor signaling in the jejunum of weanling piglets. J. Nutr. 145(1):25-31.

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Strategic Therapeutic Antibiotic Use Compared to the Challenge of Not Using

Antibiotics for Growing PigsR. Dean Boyd

Emeritus Technical Director for Hanor and Adjunct Professor of Animal Nutrition,

North Carolina State University Phone: 270-776-2536

[email protected]

Tara S. Donovan Vice President of Veterinary Management,

The Hanor Company, Enid Oklahoma

Cate E. Rush Nutrition Consultant and Research Analyst,

Savannah, Georgia

SummaryThis paper provides clarity on the challenges of not using antibiotics for therapeutic purposes, com-pared to using them strategically. A decision to not utilize medically important antibiotics in pigs, from birth (or after weaning) to harvest, should not be made without delivering on management imperatives. One must relate the increase in production cost to the ‘market opportunity’. Several groups decided to have presence in the no antibiotic ‘niche’ market, notwithstanding the possible erosion of profit. Triumph Foods conducted an experiment to serve as template for modeling the value proposition for a no therapeutic antibiotic program. In this instance, piglets did not receive antibiotics from birth to market (NAE) as compared to no antibiotics after weaning (NA-W). We also defined critical factors that must be met (e.g. PRRSv free sow herd, robust sire and sow lines, 23-25 d wean age) in order to make the NA approaches most successful. In a NA program, marginal profit is a problem for pork (and beef ), since only selected parts of the carcass are sought. Poultry have a large advantage to pork, since they can be vaccinated in the egg phase, and their life-cycle is short; however, mortality does increase and some flocks have to be removed from the NAE program. Suitable tools do not exist for total replacement of therapeutic antibiotics, to treat respiratory dis-ease. However, there are examples of antibiotic ‘alternatives’ that are auxiliary helpmates; in other words, they have not proven to replace antibiotics, but using them in combination further improves the outcome. Alternatives to enteric disease antibiotics can be more helpful, but they are not entirely satisfactory either. Zinc oxide has proven almost indispensable for weaned pigs; its use helping to reduce therapeutic treatment. Vaccine development and strict farm biosecurity also combine to re-duce therapeutic medical treatment. Complete antibiotic removal undercuts humane animal care, so they must be used for pigs that become ill. Precision rearing is more possible with pigs than with poultry, since segregation of sick pigs to medical treatment pens is logistically possible and part of normal practice. Finally, alternatives for antibiotic growth promoters are satisfactory; AGP not be-ing particularly effective with modern methods.

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Introduction This paper presents a comparison of strategic thera-peutic antibiotic use to no antibiotics ever, from birth to harvest (NAE), from the perspective of performance and financial implications. Antibiotic alternatives have not proven completely satisfactory in total replacement, but some appear to play a meaningful auxiliary role; the outcome sometimes being to improve the ability to thrive, compared to antibiotics alone. Helping food ani-mals to survive is a very different matter from the dis-cussion of alternatives to antibiotic growth promoters. In the latter case, there are some effective choices. The attraction to provide meat from animals that have never received antibiotics, follows on EU legisla-tion to remove medically important antibiotics from food animals. Some North American food corporates have made the decision to provide meat products from NAE-sourced animals (e.g. Chick-fil-A, Panera Bread, Chipotle). This has given rise to groups desiring to sat-isfy this niche with the hope of achieving a market ad-vantage. Marketing decisions were made by corporates in the Poultry sector (e.g. Perdue) to convert ‘all’ (e.g. 80-85%) of their production to that specification. Fos-ter Farms was one of the few, in the poultry sector, that elected not to follow suit on the basis of humane animal care and responsible therapeutic antibiotic stewardship. In the pork sector, Smithfield Foods made an early decision to have presence in this market (therapeutic and AGP). This was followed by Tyson and the Clem-ens Food group (NAE), and some smaller producers. At present, and for the foreseeable future (4-6 years), this pork market niche is small and driven by a small seg-ment of consumers. Food companies that have made this commitment occupy a minute share of the pork market. Product needs are easily met with present com-mitments; commoditization has been rapid. Before going directly into NAE (or NA-W) niche meat production, it is well to understand (1) what Eu-ropeans mean by ‘no antibiotics’; NAE or after weaning (NA-W); (2) how financial evaluations compare for lim-ited, therapeutic antibiotic use versus NAE and (3) what production imperatives need to be delivered on, prior to ‘eliminating’ therapeutic antibiotics for growing pigs; These 3 matters form the basis for our paper.

Understand ‘Antibiotic-free’ Program Specifications In review of many programs associated with restrict-ed or limited antibiotics, we have discovered that some are mistakenly referred to as antibiotic-free, but they are not. One such instance was in conjunction with a farm and processing plant visit in Europe, where there were

antibiotics given to every pre-weaned pig, during piglet processing. The pork product from this farm was part of an antibiotic-free market and that dose of antibiot-ics was approved and widely used by the farmers in the program. We will show, from our studies, that providing a precautionary antibiotic treatment at processing has value prior to and after weaning. Understanding the dif-ferences in specifications of the marketing program is important when evaluating the cost and performance of one program compared to another. Proof of delivery on the requirements for a NAE program is significantly more onerous that for NA-W.

Business Reality of NAE in Pig Production There are a number of road blocks to an NAE pro-gram in North America. First, the public segment is small (national, global), and market saturation was eas-ily achieved; thus evolving to a commodity product with challenged marginal return. Second, marginal cost per pig marketed, is not normally known, but it increases even on pigs derived from sow farms of high health sta-tus. The primary driver is elevated mortality, pre- and post-weaning to market. When this cost reality is jux-taposed against the demand for only selected cuts, the financial outcome is not encouraging. The third element is the most significant road block to viability – the re-spiratory pathogen PRRSv. We consider this pathogen to be the first limit to viable NAE (or NA-W) production. When the vaccine is combined with key management steps, PRRSv can be controlled, but not 100% of the time. The PRRS virus causes immune suppression with deadly, secondary pathogens (e.g. strep suis, salmonella, E. coli) arising thereafter. A key to NAE success is to control sow farm PRRSv and lateral transmission once pigs are placed in the field. Pipestone systems has had relative good success, more recently (2017 to current), with their sow farm filtration method, combined with innovative biosecurity enhancement (Dr. Barry Kerkart, pers. communication). This places them, and others like them, in a good position for NAE since, as stated, anti-biotic alternatives for the respiratory pathogens, PRRSv, Mycoplasma pneumonia and HPS, are non-existent. Extending on point number three, the number of diseases (endemic, epidemic) have increased, rather than decreased. New epidemic disease is not expected to subside with PEDv introduction into North America. Key pig growing states have also concentrated in den-sity. These are business realities that need to be consid-ered when discussing NAE. However, this point is clear – that decisions to enter the NAE niche market are usu-ally made without understanding how production cost will change.

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Value Proposition for Therapeutic Antibiotics vs NAE Triumph Foods conducted an inte-grated study to inform owner leaders, as to the cost of a NAE program, from birth to harvest (Zier-Rush et al., 2017). This data formed the template from which to model opportunity prospects, under various market conditions. The study involved comparison between conventional therapeutic medication for disease control and NAE, to harvest at approximately 290 lbs. The study was robust in that 3 robust sire lines were compared to a high performance refer-ence line (Line D). Pigs were derived from a high health sow farm; characteristic of the status required for a NAE program. Pigs in the NAE group re-ceived $2.71 in vaccine per pig, compared to $2.21 for those in the Conventional group. The cost of therapeu-tic antibiotic treatment, to treat or prevent disease was $2.78 per pig. The cost of non-antibiotic treatment (e.g. aspirin, grazix) for the NAE group was $0.85 per pig. Thus, vaccine and supportive treatment was provided to facilitate NAE viability. The test was conducted on a 4800 head, commer-cial W-F site in Iowa, which was retrofitted for research. There was a decline in full-value pigs (FVP) marketed, when antibiotic treatment was removed (feed, water, injectable). Humane care required that pigs, from either treatment, deemed to be in need of medical treatment was removed to a medical treatment pen. Their remain-ing contribution to the data involved viability. The first outcome of NAE was noted for viability to weaning at about 21 d of age. There was a 1.78% increase in prewean mortality (data not shown), when a single injection of Excede was not given, at processing, to NAE pigs (included pigs < 6 lbs at weaning). Pigs are not born into a sterile environment, so this proved that the ability to thrive benefit of precautionary antibiotic treatment

at processing. The advantage of antibiotic treatment, to prevent or treat disease, is shown in Table 1. Antibiotic treatment increased FVP, from placement to plant (94.8 vs 92.4%). This was a composite of the advantage in viability and pigs not culled to a lower value market (<230 lbs). The preponderance of mortality was due to respiratory as compared to enteric (Gut) cause, and the ability to med-ically treat pigs for respiratory challenge made a differ-ence (Figure 1). The marginal advantage to NAE strat-egy would increase further if immune stress was greater than imposed here. This may require an NAE (or NA-W program to remove a house or site from the program (Steve Pollmann, p.c.). The percent of pigs completing the NAE strategy (not removed for medical treatment) ranged from 82.3 to 77.8% for sire lines tested (Figure 2). Each of the robust lines had a higher percent of pigs that survived (data not shown), and a higher percent that re-quired no antibiotics compared to the benchmark sire. Based on our sire line comparisons, differences among sires increase as immune stress increases (data not shown). Imposing additional stress, such as pelleted diets, during elevated respiratory immune stress makes the situation even worse.

Table 1. Summary of pig viability and ability to thrive for pigs receiving no antibiotics from birth (NAE) or treated with therapeutic antibiotics (CON) to prevent or treat disease1

Variable NAE CON SEM P-ValueNo. pigs tagged at birth (EU) 2969 2953 - - Pre-wean death + rejection at wean (< 6.0 lbs), % 12.03 10.25 0.84 0.131No. pigs placed into Wean - Finish test site 2367 2234 - - Full value pigs to harvest, % placed (> 230 lbs) 92.42 94.80 0.51 0.001 Mortality, % 5.16 4.07 0.44 0.079 Cull for low weight (< 230 lbs), % 2.49 1.13 0.28 0.001Total antibiotic injections, % placed 17.61 34.95 1.50 <0.001NAE pigs marketed receiving no antibiotics, % placed 80.7 NA - -1 Pigs were pooled across sire line (Sire line x antibiotic strategy, NS) for the main effects of NAE or

Conventional treatments.

Figure 1. Proportion of mortality judged to be either respiratory, enteric (Gut) or other (known, unknown) causes. Gut and Other, P>0.33, Respiratory P=0.117.

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The primary driver of financial value was viability (Table 1), as the advantage conferred by antibiotics was negligible for growth rate (+0.02 P=0.003), feed efficiency (-0.02, NS), carcass yield (NS) and propor-tion of saleable meat per unit of carcass (NS). When all aspects of growth, viability, full-value (and cull value) price and value of saleable meat was placed against produc-tion cost, the value proposition for NAE emerged (Table 2). The relative advantage among sire lines is also shown. On the fixed-time basis that we operate under (most of the time), final weight plays a significant role in feed cost and revenue re-ceived. Under almost optimum health con-ditions, the comparative value of NAE was -$4.63 per pig placed. This estimate could be used to compute what breakeven price you would need if only 25% of the carcass cuts were saleable to an NAE market (>$16 per carcass).

Production Imperatives for Disease Resilience In order to deliver on an NAE program, every effort should be taken to deliver a healthy and disease resilient weaned pig. Negotiation to deliver on a program that commits to no antibiotics after weaning, is most desir-able. Our blue-print of management imperatives for an ‘NAE’ program, is presented in Table 3. These represent the sum of our experience, based on internal research and practice; in most cases, the scientific basis is building. The first-limiting step in an antibiotic-free option is to control sow farm PRRSv (Table 3) and lateral trans-mission after pigs are transferred to the field. The sow farm should either be PRRSv naïve, or strain presence should be the vaccine strain. A sow farm that is PRRSv stable is not acceptable, for our perspective, since this tends to be transient. PRRS virus is immunosuppressive, which opens the door to secondary bacterial pathogens (e.g. Strep suiis, Hemophilus parasuis). Resident patho-

Table 2. Summary of marginal financial differences under fixed time conditions of 164 d for NAE vs Conventional therapeutic antibiotics and for the four sire lines ($/pig placed)1.

Financial SectorComparison for Therapeutic Antibiotics Sires PooledLine A Line B Line C Line D NAE Conventional

Final weight at fixed time, lbs2 296.0 291.5 290.9 289.9 295.3 296.0Full-value pigs (>230 lbs), % 95.02 96.03 95.14 93.02 92.42 94.80Total Production Cost, $/pig placed 105.63 105.68 104.87 102.92 106.27 106.06Total Revenue including Cull Pigs, $ 154.35 151.18 150.72 147.97 149.80 154.21Net Margin, $ 48.72 45.50 45.85 45.05 43.53 48.15Marginal difference vs Control, $ 3.67 0.45 0.80 - -4.63 -1 Values extracted from the complete financial evaluation in Research Memo 2016-07. 2 Average weight of full-value and culled pigs, combined. In the NAE vs Conventional comparison – for pigs that were

removed for medication (as with Conventional pigs) their weight and financial value was not included in this analysis.

Figure 2. Comparison among sire lines for ability of their progeny to achieve market weight without antibiotic treatment (feed, water, injection). All pigs were from the NAE treatment. P-value sire line = 0.233, SEM = 1.63, Line A vs Line D, P-value = 0.052.

gens, such as mycoplasma pneumonia, exhibit greater pathogenicity with a PRRSv background. A second imperative is to utilize robust sire and sow lines. There are clear differences in sire robustness, as illustrated in Figure 2. Viability differences are also known among sow lines (internal research), but this is not as well known. We learned that segregating progeny of first-litter sows, and confining NAE to mature sow progeny is beneficial (T. Donovan, B. Meuer, p.c. 2004); the mature sow provides better protection in her colos-trum and milk than first litter females. Colostrum man-agement is likewise vital and pigs must receive a mini-mum volume (>200 g/pig; Vallet et al., et al., 2015). The best source is from the mother (Bandrick et al., 2008). Colostrum contains cellular immunity (e.g. natural kill-er cells, CD-4, CD-8) and these are specific to mom and her progeny. Cellular immune cells do not translocate to progeny blood, unless they are derived from the mother (Tuboly et al., 1988; Tuboly and Bernath, 2002). For this reason, pigs must remain with their mother for 24 h and split-sucking is advised to ensure that all pigs receive ad-equate colostrum. Antibody absorption is not mother specific, however.

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Another non-negotiable for increas-ing disease resilience is elevating wean age. Average wean age must be sufficient to keep the number of litters weaned at <21 d to less than 5%. Low wean age has proven detrimental to viability, growth (Main et al., 2004) and even lean content (Cabrera et al. 2010). The basis for this was revealed with the elegant studies of the laboratories of Moeser (Moeser, 2015; Moeser et al., 2017; Pohl et al., 2017) and Blikslager (Ziegler and Blik-slager, 2017). They showed how multiple weaning stressors, esp. early weaning (e.g 14-15 d), compromise gastrointes-tinal barrier development, and this has life-long consequences for gut health. This breach in the gut barrier has long-term consequences that include reduced viability. Thus, the mother influences her progeny, profoundly, and well beyond the colostrum phase (Odle et al., 2017). We emphasize two other facets. Geographical re-gion for pig placement and biosecurity processes must be expertly managed. Care to place pigs in a ‘pig-free’ region, however, can be foiled by intermingling of pigs, from two or more farms. Single sow farm pig flow is es-sential because death loss is lower and medical needs are less than intermingled flows (Wade et al, 2009). The biological basis for this is beginning to emerge with the study of microbial communities (enteric, tonsil, system-ic); which are remarkably diverse and environmentally specific. Sow farms appear to have a unique microflora (beneficial and pathogen).

Auxiliary Help to Antibiotics with Respiratory Challenge Antibiotic alternatives are more likely to be success-ful in preventing enteric pathogens, than for respiratory infection. In order for antibiotics to be effective against respiratory pathogens, they must be absorbed and go to the lung to be effective. In other words, getting the mol-ecule to the pathogen is what has to happen. What non-antibiotic product is known to be absorbed, and then to become lethal to the pathogen (bacteria, virus), to re-duce proliferation or enhance the immune response in the respiratory tract? A prospective alternative, whose residence remains enteric, is not expected to be effec-tive. An exception was observed by feeding spray-dried plasma to animals challenged nasally with PRRSv or In-fluenza virus. Plasma somehow caused gut active lym-phoid tissue to orchestrate changes in the lung, ahead of respiratory pathogen challenge (Campbell et al., 2016).

In our research, and from what has been reported, we are not aware of an alternative to therapeutic anti-biotics for respiratory pathogens. We have studies that show an auxiliary or additive role that an immune stim-ulant can have, when combined with therapeutic antibi-otics. Weaned pigs are the most vulnerable target, since transition from mother to dry diets and with change in residence is their most challenging ‘moment’, beyond birth. The lack of immune development is an additional risk. Strategic medication and diet format (composi-tion, budget) can minimize the challenge during this immunologically vulnerable time. Pathogenic disease-stressors cause the pig to utilize the innate immune system, to the extent that it has de-veloped. Medications are a means to reduce pathogen exposure and this partially compensates for an under-

Table 3. Management imperatives that are not negotiable for No Antibiotic pig production.

• Sow farm free of the disease pathogen, PRRSv• Robust sire line• Robust sow line(s)• Immature first-litter female progeny are immune inferior to

mature progeny (cattle, sows)• Minimum Colostrum intake from maternal sow• Minimum 23-25 day wean age (<5% under 21 days age)• Strategic antibiotic injection at processing (not born into

sterile environment)• Pigs placed in single sow farm flows (not intermingled)• Place weaned pigs in low pig density area (pathogen

plumes, cause lateral infections)• All-in All-out site management to reduce pathogen concen-

tration• Female pig flows thrive better than males• Vaccines field validated for effectiveness• Transition Diets stress minimum (composition, budget deliv-

ered on)• Pigs requiring antibiotic medication removed to medical

pens (advantage to poultry)

Figure 3. Comparison of therapeutic antibiotics (MED) to no antibiotics after weaning (NO MED) and for an algae source of 1-3 β-glucan (Proglena), alone or in combination with therapeutic antibiotics

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developed immune response. Our lat-est study (Dion and Boyd, 2018) illus-trates this facet and the format we use to test proposed antibiotic alternative challengers; in conditions of low and high respiratory immune stress (e.g. PRRSv stable, unstable sow farm). This study shows that a specific algae source of 1-3 β-glucan (>90% of β-glucan is in the 1-3 configuration) added to the benefit observed with therapeutic anti-biotics alone, in the face of a respiratory challenge. In the absence of antibiotics, it provided no measurable improve-ment, thereby showing the value of antibiotics in combatting respiratory assault, and the additive nature of im-proving the response by an immature immune system. Earlier, we observed immune enabling with a yeast source of 1-3 β-glucan (Cenzone Yeasture). The objective of the study was to compare our thera-peutic antibiotic program (Pulmotil, CTC/Denagaard), for treating weaned pigs derived from a sow farm with active PRRSv virus (unstable), to a non-antibiotic chal-lenger (algae source 1-3 β-glucan, proglena), for growth, viability and ability to thrive (indicated by medical treat-ment, low weight culls). A negative control (no antibiot-ics) group was compared to the antibiotic control. The

Table 4. Growth and viability comparison of therapeutic antibiotics (MED) to no antibiotics after weaning (NO MED) and for an algae source of 1-3 β-glucan (Proglena), alone or in combination with therapeutic antibiotics1

ItemPOS CTR

MEDNEG CTR NO MED

NEG CTR + Proglena

POS CTR + Proglena SEM TRT 1v2

No. Pens 19 20 20 19 - - -No. Pigs 418 440 440 418 - - -Initial Weight, lbs 15.7 15.6 15.7 15.6 0.65 0.641 0.336Period 1, 0-20 d2

End Weight, lbs 29.5 27.6 27.6 29.8 0.90 <0.001 <0.001Gain, lbs 13.7 12.0 11.9 14.2 0.38 <0.001 <0.001ADG, lbs/d 0.69 0.60 0.60 0.71 0.02 <0.001 <0.001ADFI, lbs/d 0.80 0.75 0.75 0.81 0.02 0.007 0.023FCR, feed/gain ratio 1.16 1.25 1.25 1.15 0.02 <0.001 <0.001Nursery Period, 0-39 d2

End Weight, lbs 49.5 45.5 46.0 50.4 1.37 <0.001 <0.001Gain, lbs 33.8 30.4 30.4 34.8 0.85 <0.001 <0.001ADG, lbs/d 0.87 0.78 0.78 0.89 0.02 <0.001 <0.001ADFI, lbs/d 1.17 1.08 1.07 1.18 0.03 <0.001 <0.001FCR, feed/gain ratio 1.35 1.40 1.38 1.34 0.01 <0.001 <0.001

Full-Value Pigs, % 97.0 95.7 93.9 98.8 1.24 0.002 0.427Average injections per pen 1.3 10.4 9.9 2.6 0.83 <0.001 <0.0011 Initial age = 22.8 days, ranged from 19 to 27 days.2 Normal performance traits did not account for dead pig weights, and included: ADG = (avg. pig weight end

– avg. pig weight initial)/ (days on the period); ADFI= (feed delivered- feed weighed back at the end of the period)/ (pig days on the period); FCR= ADFI/ADG. FVP is defined as the percentage of pigs that weighed > 24.2 lbs at the end of the trial (<4 standard deviations from the mean, which is possible where diet stress exists resulting in a skewed left tail of the distribution).

modality of proglena was to facilitate earlier develop-ment of the adaptive immune system (Dalmo and Bog-wald, 2008). This source of the 1-3 β-glucan (Euglena gracilis) was of particular interest because the algae pro-vided survival protection against influenza virus infec-tion in mice (Nakashima et al., 2017). The study involved 2,156 weaned pigs (22.8 d avg age; range, 19 to 27 d) that were PRRSv (1-7-4) positive at weaning. Pigs were removed to medical pens, as re-quired, but remained on respective diet treatments. The nursery remained positive for PEDv and porcine delta-

Figure 4. Comparison of therapeutic antibiotics (MED) to no antibiotics after weaning (NO MED) and an algae source of 1 -3 β-glucan (Proglena), alone or in combination with therapeutic antibiotics using Pen ADG and FCR. This is the multiple of viability (retention) and individual pig ADG and FCR.

Figure 4. Comparison of therapeutic antibiotics (MED) to no antibiotics after weaning (NO MED) and an algae source of 1 -3 β-glucan (Proglena), alone or in combination with therapeutic antibiotics using Pen ADG and FCR. This is the multiple of viability (retention) and individual pig ADG and FCR.

Figure 5. Financial evaluation for marginal return over cost for therapeutic antibiotics (MED) to no antibiotics after weaning (NO MED) and an algae source of 1-3 β-glucan (Proglena), alone or in combination with therapeutic antibiotics.

17.2915.60 15.33

18.64

1.431.51 1.52

1.37

1.201.251.301.351.401.451.501.551.60

0.50

5.50

10.50

15.50

20.50

POS CTRMED

NEG CTRNO MED

NEG CTR +Proglena

POS CTR +Proglena

0‐39

d FCR

, Feed : G

ain

0‐39

d ADG

, Lbs/d/pen

Nursery Pen Performance (15‐48 lbs)Pen ADG, lbs/d Pen Feed:Gain ratio

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Figure 5. Financial evaluation for marginal return over cost for therapeutic an-tibiotics (MED) to no antibiotics after weaning (NO MED) and an algae source of 1-3 β-glucan (Proglena), alone or in combination with therapeutic antibiotics.

corona virus throughout the study. Pigs were considered as under moderate im-mune stress. Performance is summarized in Ta-ble 4. The primary driver of the value proposition is improved FVP, and this was compromised by not using antibi-otics (97.0 vs 95.7% for not medicated; treatments, P<0.002). Proglena alone did not reduce mortality (Figure 3), but when combined with antibiotics, mor-tality tended to decline compared to antibiotics alone (0.97 vs 2.51%); further, FVP was numerically improved (98.8 vs 97.0%, P=0.29); the latter would require about 100 EU pens/treatment to prove statistically (SD=4.8) at the 0.05 level of significance. The number of Baytril in-jections (doses per pen) was 8-fold higher for No MED as compared to MED pigs (P<0.05). The impact of MED and MED plus immune facilitation, on the ability to thrive, as compared to No MED was reflected in im-proved ADG and FCR (Table 4). When death rate is dif-ferent among treatments, the multiple of retained pigs and growth rate is a better illustration of the treatment effect – pen ADG and pen FCR (Figure 4). The financial outcome presented in Figure 5 con-sidered gain value, but it also accounted for the value of dead weaned pigs, which are not free ($30-36/weaned pig cost). Treatments that elevate death loss during the nursery phase include (a) no medication and (b) cheap diet composition (or low budget). This might also be applied to reducing the dose of a life-saving vaccine to reduce cost, if that decision results in greater death. Failure to use MED resulted in a negative return over investment (ROI) of -$3.33/pig, but the combination of MED and Proglena β-glucan increased the marginal ROI to $1.12 per pig out. Thus, MED is the most effec-tive means to minimize death from respiratory disease, that we have tested in this critical period; but facilita-tion of the adaptive immune system, is a comparatively cheap auxiliary tool to further improve viability and FCR. This would be more difficult to prove under con-ditions of high health.

Disease Resilience, Health Promotion and AGP Alternatives Global respiratory disease threat requires the use of medically important antibiotics to survive. Diseases include PRRSv, influenza, mycoplasma pneumonia, he-mophillus parasuis and their mutation variants. How-ever, the global pathogen library is even more ominous

than this. Antibiotics are a powerful pathogen control-ling or ameliorating technology. Failure to control respi-ratory disease leads to secondary, bacterial challenges (respiratory, enteric), increased mortality and reduced sustainability. On the other hand, growth promoting antibiotics (AGP) are not essential to pig production. More effective alternatives are emerging and these tend to improve gut barrier resilience and reduce enteric pathogen load through various mechanisms. Given the disease threats that are before the Ameri-can Pig Industry, and with a mind toward reducing de-pendence on therapeutic antibiotics, or elimination in the case of a No Antibiotics program, Table 5 provides example tactics (classes) that are deemed sufficiently ef-fective to be noted. The framework presents examples of alternative antibiotic growth promoters (AGP) as well as those that significantly reduce pathogens (lethal-ity, proliferation, attachment); pathogens that would otherwise increase mortality. We specifically avoided listing the ‘kitchen sink’ of alternative AGP and refer readers to reviews by Liu et al. (2018), Pluske (2013) and Heo et al. (2012) and to footnote 1 of Table 5.

Modest Look Forward – Medically Important Antibiotics in Food Animals We do not expect antibiotics, which are medically important to food animals, to be eliminated in our working life-time by American legislative mandate. We believe that there will be more pressure placed on docu-mentation to trace outcomes relative to stated purpose for use. This is what Europe has evolved to; did it work or not work ? Greater regulation around the use of ther-apeutic antibiotics, specifically critically important for human use, is expected.

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The AVMA established ‘Core Principles of Antimi-crobial Stewardship’ that we ascribe to (https://www.avma.org/KB/Policies/Pages/Antimicrobial-Steward-ship-Definition-and-Core-Principles.aspx). Central to the 5 principles, is using an evidence-based approach in making decisions to use antimicrobial drugs and then continually evaluating outcomes of the therapy. Technologies such as the PRRSv resistant pig and better vaccine technologies will be key prevention mechanisms for disease. A recent decision by the EU court, to categorize gene deletion as creating a GMO (PRRSv crispr technology) is a set-back, but this deci-sion may not be widely accepted in key global markets. There are also other mechanisms of selecting for re-

Table 5. Example tactics deemed sufficiently effective in promoting immunity or growth1.

Category Class or Aspect ExplanationProduction technique Sow farm is foundation Air filtration, Biosecurity, Location, Farm size

Genetic lines Viability under disease stress is very differentColostrum, mother progeny Immune cells, antibodies; minimum 200 g/pigSow Milk Bioactive moleculesWean age, >23 d Early weaning (14-16 d), immune compromise

Immune enabling Vaccines Strong therapeutic antibiotic complimentWean pig diets 1-3 ‘Cheap’ composition, Inadequate budget causes

Pathogen proliferation, Barrier disruption1-3 β-glucan (algae, yeast) Facilitates Adaptive immune elementSpecific Nutrients E, D (esp. 25-OH D3), C, Zn, Cu, Arginine, 18:2

Immune tempering – Aspirin, salicylates, dexamet. Tempers excessive lethal inflammation(Inflammation hyper-response) Animal plasma Reduce inflammation

Soybeans Reduce inflammation (e.g. Isoflavones)Fish meal Reduce inflammation

Respiratory auxiliary help2 Animal plasma Oral consumption, systemic immune response1-3 β-glucan (algae, yeast) Algae beneficial to Influenza challenged mice

Resp. antibiotic alternative3 Unaware of proven alternative See Table 4, Figures 3-4Enteric health, growth Zinc Oxide Proven E. Coli controlpromotion (See Pluske 2013) Copper salts Pathogen control, growth promotion

Egg Immunoglobulins Targeted Pathogen controlEnzyme Xylanase Improve viability via favorable microbiota shiftBotanicals Pathogen control (Grazix, Carvacrol, Thymol)Medium-chain fatty acids Pathogen control (6, 8, 10 carbons)Organic acids Pathogen control, growth promotion

Emerging tactics – pathogen Gene editing Crispr technology, PRRSv attachment inabilityControl (respiratory, enteric) Epidermal growth factor Oral EGF ameliorates diarrhea, epithelia repair4

Bacteriocins Colicin, Anti-microbial peptidesBacteriophage viruses Replicate in host to destroy pathogen

OTHER: Metabolic Modifiers5 Ractopamine β-agonist stimulation of protein synthesisand Immuno-Castration Anti-GNRF injection allows intact benefit for timeDigestion end-products5 Ionophore Narasin Binds gram (+) bacteria for hypertonic death

Propionate : Acetate + Butyrate ratio increases1 List of enteric alternatives for antibiotic growth promotion (proactive pathogen disruption) is long. Classes include probiotics, prebiotic

fermentable substrates, specifically stressed yeast to generate internal stress-resistant biomolecules, dietary carbohydrate modification (less soluble NDF, less total NDF) and a longer list of botanical products (than named above) including capucins (antioxidant reduction). Interferons are likewise noted as these naturally occurring molecules interfere with virus reproduction.

2 Auxiliary helps to antibiotics when respiratory system becomes infected.3 Antibiotic alternatives to antibiotic treatment of respiratory infection (e.g. PRRSv, Influenza, Hemophilus parasuis).4 Basis for this prospective technology is presented by Zijltra et al. 1994 (repair epithelia, villi after rotavirus infection of piglets) and

Yonghua et al. 2016 (Soya-sourced r-EGF found to be as bio-efficacious as human EGF in mechanism involved in preventing necrotizing enterocolitis). SOYA presented as a prospective biopharma generator of EGF, which promotes epithelial growth, proliferation, cytokine response tempering, maximizing digestive enzyme secretions, decreases cell death and IgA generation to ameliorate gut allergens.

5 This class is intended to bring attention to example components whose mechanism is not associated with Pathogen control, per se; rather to change Metabolism or Digestion end products. These are examples of a growing list.

duced disease responsiveness in the pig (molecular typ-ing and mapping) and on disease agents (e.g. modified live pathogen vaccines selected against virulence).

ReferencesBandrick, M., M. Pieters, C. Pijoan, and T.W. Molitor.

2008. Passive transfer of maternal mycoplasma hy-opneumoniae-specific cellular immunity to piglets. Clinical and Vaccine Immunology. Vol 15:3: 540-543.

Cabrera, R., R.D. Boyd, S. Jungst, E. Wilson, M. John-ston, J. Vignes, and J. Odle. 2010. Impact of lacta-tion length and piglet weaning weight on long-term growth and viability of progeny. J. Anim. Sci. 88: 2265-2276.

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Campbell, J., J.Polo, and J.Crenshaw. 2016. Orally fed spray dried plasma modulated the immune re-sponse during respiratory challenges: A review. J. Anim. Sci. 94:45-47.

Díaz, I., C. Lorca, I. Galindo, J. Campbell, I. Barranco, L. Kuzemtseva, I. M. Rodriguez-Gomez, J. Crenshaw, L. Russell, and J. Polo., J. Pujols. 2010. Potential posi-tive effect of commercial spray-dried porcine plas-ma on pigs challenges with PRRS virus. Proc. 21st IPVS Congress, 21:560.

Dalmo, R. and J. Bogwald. 2008. B-Glucans as conduc-tors of immune symphonies. Fish and Shellfish Im-munology 25:384-396.

Dion, K and R.D. Boyd. 2018. Comparison of two non-antibiotic health promotion regimens to current Hanor antibiotic program for growth, viability and FVP during the nursery period (13-50 lbs). Hanor Technical Research Memo H 2017-16 (available from authors, Boyd, Donovan).

Heo, J., F. Opapeju, J. Pluske, J. Kim, D. Hampson and C. Nyachoti. 2012. Gastrointestinal health and func-tion in weaned pigs: A review of feeding strategies to control post-weaning diarrhoea without using in-feed antimicrobial compounds. J. Anim. Physio. Anim. Nutr. 97:207-237.

Liu, Y., C Espinosa, J. Abelilla, G. Casas, L. Lagos, S. Lee, W. Kwon, J. Mathai, D. Navarro, N. Jaworski, H. Stein. 2018. Non-antibiotic feed additives in diets for pigs: A review. Animal Nutrition (In Press).

Main, R.G., S.S. Dritz, M.D. Tokach, R.D. Goodband and J.L. Nelssen. 2004. Increasing weaning age im-proves pig performance in a multisite production system. J. Anim. Sci. 82:1499-1507.

Moeser, A. 2015. Gut barrier function in pigs: influence of stress, management, and nutrition. St Paul (MN): Minnesota Nutrition Conference.

Moeser, A., C. Pohl, and M. Rajput. 2017. Weaning stress and gastrointestinal barrier development: Im-plications for lifelong gut health in pigs. Animal Nu-trition. 3:313-321.

Nakashima, A., K. Suzuki, Y. Asayama, M. Konno, K. Saito, N. Yamazaki, and H. Takimoto. 2017. Oral ad-ministration of Euglena gracilis Z and its carbohy-drate storage substance provides survival protection against influenza virus infection in mice. Biochemi-cal and Biophysical Research Communications. 494:379-383.

Odle, J., S. Jacobi, R.D. Boyd, D, Bauman, R. Anthony, F. Bazer, A. Lock, and A. Serazin. 2017. The potential impact of animal science research on global mater-nal and child nutrition and health: A landscape re-view. Adv. Nutr. 8:362-381.

Pluske, J.R. 2013. Feed and feed additives-related as-pects of gut health and development in weanling pigs. J. Anim. Sci. Biotech. 4:1-7.

Pohl, C., J. Medland, E. Mackey, L. Edwards, K. Bagley, M. DeWilde, K. Williams, and A. Moeser. 2017. Ear-ly weaning stress induces chronic functional diar-rhea, intestinal barrier defects, and increased mast cell activity in a porcine model of early life adversity. Neurogastroenterol Motil. 29:e13118.

Tuboly, S., S. Bernath, R. Glavits, and I. Medveczky. 1988. Intestinal absorption of colostral lymphoid cells in newborn piglets. Veterinary Immunoloty and Immunopathology. 20:75-85.

Tuboly, S. and S. Bernath. 2002. Intestinal absorption of colostral lymphoid cells in newborn animals. Adv Exp Med Biol. 503:107-114.

Vallet, J., J. Miles, L. Rempel, D. Nonneman, C. Lents. 2015. Relationships between day one piglet serum immunoglobulin immunocrit and subsequent growth, puberty attainment, litter size and lactation performance. J. Anim. Sci. 93:2722-2729.

Wade, D., T. Donovan, J. Moody, R. Rott. 2009. Single-source versus multi-source pig flows. Allen D. Le-man Swine Conference. 2009:128-129. Retrieved from the University of Minnesota Digital Conservan-cy, http://hdl.handle.net/11299/139776

Yonghua, H., M.A. Schmidt, C. Erwin, J.Guo, R. Sun, K. Pendarvis, B.W. Warner, E. Herman. 2016. Transgenic soybean production of bioactive hu-man epidermal growth factor (EGF). PLoS ONE 11(6):e0157034.

Ziegler A, and A. Blikslager. 2017. Impaired intestinal barrier function and relapsing digestive disease: Les-sons from a porcine model of early life stress. Neuro-gastroenterol Motil. 29:e13216.

Zier-Rush, C., M. Tillman, M. Rauen, W. Barrett, C. Groom, A. Elsbernd, D. McAlister, E. Arkfeld, J. Moody, T. Donovan, K. Dion, C. Minor, M. McCul-ley and R.D. Boyd. 2017. Comparison of the prog-eny of 4 sire lines for integrated profit using either conventional antibiotic or no antibiotic treatment from birth to market under high health (14-293 lbs). Hanor Research Memo H 2016-07 (available from authors, Boyd, Donovan).

Zijlstra, R.T, J. Odle, W.F. Hall, B.W. Petschow, H.B. Gel-berg and R.E. Litov. 1994. Effect of orally adminis-tered epidermal growth factor on intestinal recovery of neonatal pigs infected with rotavirus. J. Pediatric Gastroen. Nutrition 19:382-390.

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2017 Conference

Bob Easter, University of Illinois, Keynote speaker

Brian Richert, Purdue University, Speaker

Charles Woloshuk, Purdue University, Speaker

Kirk Klasing, University of California, Davis, Speaker

Hans Stein, University of Illinois, Speaker

Lowell Randel, The Randel Group, Alexandria, VA, Speaker

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Discussing Brian Richert’s presentation.

2017 Conference

Charles Zila and Sara Larsson, DuPont Pioneer, Windfall, IN, Speakers

Gretchen Hill, Michigan State University, Speaker Dennis Liptrap, Ralco Nutrition, Moderator

Pioneer’s graph showing yield of corn, wheat, and soybeans from 1900 to 2016.

Scott Radcliffe, Purdue University, Videographer

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2017 Conference

Over 100 attendees listening to a presentation.

Pork loin luncheon for the attendees. Dining in the conference room.

Four screens showed slides of speakers.

Lots of visiting occurs during the breaks.Attendee enjoying the meal.


Number of Years of SponsorshipNo. of Years Company

18 AlltechUnited Animal Health (previously JBS United)

17 Elanco Animal Health Hubbard FeedsProvimi North America (previously Akey) Purina Animal Nutrition (previously Land

O’Lakes/Purina Mills)

16 PIC North America Zinpro Corporation

15 Ajinomoto Animal Nutrition (previously Ajinomoto Heartland)

APC CompanyDSM Nutritional ProductsEvonik-Degussa CorporationInternational Ingredient CorporationNovus International

14 Agri-KingBASF CorporationFats and Proteins Research FoundationPrince Agri Products

13 Cooper FarmsDuPont (previously Danisco Animal Nutrition)National Pork BoardRalco NutritionZoetis (previously Pfizer Animal Health,

Alpharma)

12 ADM Animal NutritionChr. Hansen Animal Health and NutritionDistributors ProcessingDarling Ingredients (previously Griffin

Industries)Kemin

11 Kent Nutrition GroupPhibro Animal Health

No. of Years Company

9 Pioneer Hi-Bred InternationalPOET NutritionThe Maschhoffs

8 Lallemand Animal NutritionMicronutrientsNovartis Animal HealthStuart Products

7 AB Vista Feed IngredientsMonsantoVita Plus Corporation

6 Diamond V MillsKalmbach FeedsNutriQuest

5 King Techina GroupNewsham Choice GeneticsNutriad

4 Cargill Animal NutritionHamlet ProteinPharmgate Animal Health

3 CJ AmericaPancosma

2 CHSMurphy-BrownMycogen SeedsPfizer

1 ChemGenGladwin A. Read Co.NRCS Conservation Innovation GrantNutrafermaStandard Nutrition ServicesVi-Cor Animal Health and Nutrition

New sponsors this year:CSA Animal NutritionNorel Animal NutritionPhileo Lesaffre Animal Care

18th annual midwest swine nutrition conference proceedings€¦ · cj america cooper farms csa...

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